T cells require a signal induced by the engagement of the T cell receptor and a “co‑stimulatory” signal(s) through distinct T cell surface molecules for optimal T cell expansion and activation. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co‑stimulatory molecules found on the surface of T cells. Members of the B7 superfamily include B7-1 (CD80), B7-2 (CD86), B7‑H1 (PD-L1), B7-H2 (B7RP-1), B7-H3, and PD-L2 (1). B7-H3 is expressed at very high levels in immature dendritic cells at moderate levels on mature dendritic cells, LPS stimulated immature dendritic cells and LPS stimulated monocytes, and at low levels on resting monocytes. B7-H3 binds to activated T cells via an as-of-yet identified receptor. B7-H3 co-stimulates proliferation of T cells and interferon-gamma (IFN-gamma ) production and enhances the induction of cytotoxic T cells. B7-H3 shares 20‑27% amino acid (aa) identity with other B7 family members (2). Murine B7-H3 is a 259 aa protein containing an extracellular domain, a transmembrane domain and a cytoplasmic domain. Mouse and human B7-H3 share 87% aa identity (3).
Key Product Details
Species Reactivity
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Applications
Validated:
Cited:
Label
Antibody Source
Product Specifications
Immunogen
Val29-Phe244
Accession # Q8VE98
Specificity
Clonality
Host
Isotype
Endotoxin Level
Scientific Data Images for Mouse B7‑H3 Antibody
Detection of Mouse B7‑H3 by Western Blot.
Western blot shows lysates of MEF mouse embryonic feeder cells, P19 mouse embryonal carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, C2C12 mouse myoblast cell line, and 3T3-L1 mouse embryonic fibroblast adipose-like cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 45-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Proliferation Induced by B7‑H3 and Neutralization by Mouse B7‑H3 Antibody.
Recombinant Mouse B7-H3 enhances proliferation in mouse CD3+T cells in the presence of 100 ng/mL Hamster Anti-Mouse CD3e Monoclonal Antibody (Catalog # MAB484) in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse B7-H3 (2 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397). The ND50 is typically 1.5-7.5 µg/mL.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Mouse B7‑H3 Antibody
Western Blot
Sample: MEF mouse embryonic feeder cells, P19 mouse embryonal carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, C2C12 mouse myoblast cell line, and 3T3‑L1 mouse embryonic fibroblast adipose-like cell line
Neutralization
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: B7-H3
References
- Coyle, A.J. and J.-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
- Chapoval, A.I. et al. (2001) Nature Immunol. 2:269.
- Sun, M. et al. (2002) J. Immunol. 168:6294.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional B7-H3 Products
Product Documents for Mouse B7‑H3 Antibody
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Product Specific Notices for Mouse B7‑H3 Antibody
For research use only
Citations for Mouse B7‑H3 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
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