Human SGK1 Antibody

Detection of Human SGK1 by Western Blot. Western blot shows lysates of CHO Chinese hamster ovary cell line non-transfected and transfected with human SGK1. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human SGK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3200) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for SGK1 was detected at approximately 54 kDa (as indicated). For additional reference, recombinant human SGK1, SGK2, and SGK3 were included. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of SKG1 in Human Kidney. SKG1 was detected in paraffin-embedded sections of human kidney using Rabbit Anti-Human SGK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3200) at 3 μg/mL for 1 hour at room temperature followed by incubation for 30 minutes at room temperature with Anti-Rabbit IgG VisUCyte HRP Polymer Antibody (VC003). Tissue was stained with DAB (brown color) and counterstained with hematoxylin (blue color).

Detection of Human SGK1 by Western Blot SGK1 inhibits Tas from transactivating PFV LTR and IP promoters. Levels of integrated proviral DNA were measured using semiquantitative PCR (A) and real-time PCR (B). (C) HT1080 cells (1 × 105) were transfected with LTR-Luc (0.05 μg) or IP-Luc (0.025 μg), combined with 3.1-Tas and empty vector or SGK1. At the same time, pCMV-beta -gal (0.05 μg) was transfected to normalize the transfection efficiency. At 48 h posttransfection, luciferase activities were measured and corrected by beta -gal catalytic activities. The remaining cell lysates were used for Western blotting. (D) HT1080-siControl and HT1080-siSGK1 cells (1 × 105) were transfected with LTR-Luc (0.05 μg) or IP-Luc (0.025 μg) and 3.1-Tas. At the same time, pCMV-beta -gal (0.05 μg) was transfected to normalize transfection efficiency. At 48 h posttransfection, luciferase activities were measured and corrected by beta -gal catalytic activities. The remaining cell lysates were used for Western blotting. Data are expressed as the means ± standard deviations. Data are representative of three independent experiments. One-way ANOVA was used to perform the statistical test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ns for P > 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35438526), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human SGK1 by Western Blot SGK1 inhibits PFV replication. (A to C) HT1080-Control and HT1080 stable expression cells (1 × 105) were infected with PFV (MOI = 0.6). At 48 h postinfection, 600 μL of the supernatants (A) or 1/10 infected HT1080 cell lines (B) were incubated with PFVL cells (1 × 105), the luciferase activity was measured 48 h later, and the rest of the infected cells were lysed for Western blotting (C). (D to F) HT1080-siControl and HT1080-siSGK1 cells (1 × 105) were infected with PFV (MOI = 0.6). At 24 h postinfection, 600 μL of the supernatants (D) or 1/10 infected HT1080 cells (E) were incubated with PFVL cells (1 × 105), and the luciferase activity was measured 48 h later. (F) The rest of the infected cells were lysed for Western blotting. (G to I) PFV (MOI = 0.6) infected Control and sg3 cell lines (1 × 105). At 48 h postinfection, 600 μL of the supernatants (G) or 1/10 infected HT1080 cells (H) were incubated with PFVL cells (1 × 105), and the luciferase activity was measured 48 h later. (I) The remaining infected cells were lysed for Western blotting. Data are expressed as the means ± standard deviations. Data are representative of three independent experiments. One-way ANOVA was used to perform the statistical test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35438526), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human SGK1 by Western Blot SGK1 inhibits Tas from transactivating PFV LTR and IP promoters. Levels of integrated proviral DNA were measured using semiquantitative PCR (A) and real-time PCR (B). (C) HT1080 cells (1 × 105) were transfected with LTR-Luc (0.05 μg) or IP-Luc (0.025 μg), combined with 3.1-Tas and empty vector or SGK1. At the same time, pCMV-beta -gal (0.05 μg) was transfected to normalize the transfection efficiency. At 48 h posttransfection, luciferase activities were measured and corrected by beta -gal catalytic activities. The remaining cell lysates were used for Western blotting. (D) HT1080-siControl and HT1080-siSGK1 cells (1 × 105) were transfected with LTR-Luc (0.05 μg) or IP-Luc (0.025 μg) and 3.1-Tas. At the same time, pCMV-beta -gal (0.05 μg) was transfected to normalize transfection efficiency. At 48 h posttransfection, luciferase activities were measured and corrected by beta -gal catalytic activities. The remaining cell lysates were used for Western blotting. Data are expressed as the means ± standard deviations. Data are representative of three independent experiments. One-way ANOVA was used to perform the statistical test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ns for P > 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35438526), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human SGK1 by Western Blot SGK1 inhibits the transactivation function of Tas in a kinase-independent manner. (A to C) HEK293T cells (2 × 105) were co-transfected with pcPFV (0.5 μg) and empty vector, S422D, SGK1, or K127N (0.5 μg). At 48 h posttransfection, 600 μL of the supernatants (A) or 1/20 transfected cells (B) were incubated with PFVL cells (1 × 105), the luciferase activity was measured 48 h later. (C) The rest of transfected cells were lysed for Western blotting. (D, E) HEK293T cells (2 × 105) were transfected with LTR-Luc (0.025 μg) (D) or IP-Luc (0.01 μg) (E), combined with 3.1-Tas (0.1 μg) and empty vector or SGK1, SGK1-S422D, and SGK1-K127N (0.3 μg). At the same time, pCMV-beta -gal (0.025 μg) was transfected to normalize the transfection efficiency. At 48 h posttransfection, luciferase activities were measured and corrected by beta -gal catalytic activities. Remaining cell lysates were used for Western blotting. Data are expressed as the means ± standard deviations. Data are representative of three independent experiments. One-way ANOVA was used to perform the statistical test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; and ns for P > 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35438526), licensed under a CC-BY license. Not internally tested by R&D Systems.