Human IL-4I1 Antibody

Detection of Human IL‑4I1 by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS for 3 hours. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human IL-4I1 Monoclonal Antibody (Catalog # MAB5684) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for IL-4I1 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL‑4I1 in HDLM‑2 Human Cell Line. IL-4I1 was detected in immersion fixed HDLM-2 human Hodgkin's lymphoma cell line using Rat Anti-Human IL-4I1 Monoclonal Antibody (Catalog # MAB5684) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in lysosomes. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

IL‑4I1 in Human B Cell Lymphoma. IL-4I1 was detected in immersion fixed paraffin-embedded sections of human B cell lymphoma using Rat Anti-Human IL-4I1 Monoclonal Antibody (Catalog # MAB5684) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC005). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of IL-4I1 by Western Blot IFN-gamma and TNF-alpha increase the expression of IL4I1 in MuSCs through NF-kappa B and STAT6 pathway.A Volcano plot of differentially expressed genes in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. B Volcano plot of differentially expressed genes in MSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. C The mRNA expression of IL4I1 in MSCs and MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was assayed by qRT-PCR. D The concentration of IL4I1 in the supernatants of MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was measured by ELISA. E The protein expression of IL4I1 and beta -ACTIN (loading control) in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was determined by western blotting. F The mRNA and protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of AS1517499 were respectively assayed by qRT-PCR and western blotting. G The protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of BAY117082 were assayed by western blotting. Data were shown as means ± SEM. Data were representative of three experiments with similar results. For two-group comparison, statistical analysis was performed by Student’s t test. ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-4I1 by Western Blot IL4I1 mediates the therapeutic effect of MuSCs on ALI.A The therapeutic strategy of MuSCs in the LPS-induced ALI model. Mice were treated with 2 mg/kg LPS through endotracheal infusion. 1 h later, Ctrl-shRNA MuSCs or IL4I1-shRNA MuSCs (5 × 105) pretreated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h were intravenously injected into mice. Then, all experimental mice were euthanized after 23 h, and the lung samples were collected for further processing. B The efficiency of IL4I1 knockdown measured by western blotting analysis. C The total amount of IL4I1 protein in the lung tissue homogenates of ALI mice was determined by ELISA (PBS: n = 3, LPS: n = 3, LPS+Ctrl-shRNA MuSCs: n = 3, LPS + IL4I1-shRNA MuSCs: n = 3). D The expression levels of IL-6 mRNA (left) and protein (right) in the lung tissue homogenates of ALI mice were respectively determined by qRT-PCR and ELISA. E Lung tissues of mice with various treatments were fixed for H&E staining. Yellow arrowhead, area of widespread septal thickening with increased air-space cellularity and exudation and enhanced interstitial immune cell infiltration in the damaged lungs of ALI mice. Scale bars, 250 μm. F The expression levels of chemokines in the lung tissue homogenates of ALI mice were determined by qRT-PCR. G Lung tissues of mice with various treatments were stained with CXCL1 antibody. Scale bars, 250 μm (PBS: n = 3, LPS: n = 4, LPS+Ctrl-shRNA MuSCs: n = 5, LPS + IL4I1-shRNA MuSCs: n = 5). Data were shown as means ± SEM. Data were representative of three experiments with similar results. For multiple group comparison, statistical analysis was performed by one-way ANOVA 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/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-4I1 by Western Blot IFN-gamma and TNF-alpha increase the expression of IL4I1 in MuSCs through NF-kappa B and STAT6 pathway.A Volcano plot of differentially expressed genes in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. B Volcano plot of differentially expressed genes in MSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. C The mRNA expression of IL4I1 in MSCs and MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was assayed by qRT-PCR. D The concentration of IL4I1 in the supernatants of MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was measured by ELISA. E The protein expression of IL4I1 and beta -ACTIN (loading control) in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was determined by western blotting. F The mRNA and protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of AS1517499 were respectively assayed by qRT-PCR and western blotting. G The protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of BAY117082 were assayed by western blotting. Data were shown as means ± SEM. Data were representative of three experiments with similar results. For two-group comparison, statistical analysis was performed by Student’s t test. ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-4I1 by Western Blot IFN-gamma and TNF-alpha increase the expression of IL4I1 in MuSCs through NF-kappa B and STAT6 pathway.A Volcano plot of differentially expressed genes in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. B Volcano plot of differentially expressed genes in MSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. C The mRNA expression of IL4I1 in MSCs and MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was assayed by qRT-PCR. D The concentration of IL4I1 in the supernatants of MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was measured by ELISA. E The protein expression of IL4I1 and beta -ACTIN (loading control) in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was determined by western blotting. F The mRNA and protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of AS1517499 were respectively assayed by qRT-PCR and western blotting. G The protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of BAY117082 were assayed by western blotting. Data were shown as means ± SEM. Data were representative of three experiments with similar results. For two-group comparison, statistical analysis was performed by Student’s t test. ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-4I1 by Western Blot IL4I1 mediates the therapeutic effect of MuSCs on ALI.A The therapeutic strategy of MuSCs in the LPS-induced ALI model. Mice were treated with 2 mg/kg LPS through endotracheal infusion. 1 h later, Ctrl-shRNA MuSCs or IL4I1-shRNA MuSCs (5 × 105) pretreated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h were intravenously injected into mice. Then, all experimental mice were euthanized after 23 h, and the lung samples were collected for further processing. B The efficiency of IL4I1 knockdown measured by western blotting analysis. C The total amount of IL4I1 protein in the lung tissue homogenates of ALI mice was determined by ELISA (PBS: n = 3, LPS: n = 3, LPS+Ctrl-shRNA MuSCs: n = 3, LPS + IL4I1-shRNA MuSCs: n = 3). D The expression levels of IL-6 mRNA (left) and protein (right) in the lung tissue homogenates of ALI mice were respectively determined by qRT-PCR and ELISA. E Lung tissues of mice with various treatments were fixed for H&E staining. Yellow arrowhead, area of widespread septal thickening with increased air-space cellularity and exudation and enhanced interstitial immune cell infiltration in the damaged lungs of ALI mice. Scale bars, 250 μm. F The expression levels of chemokines in the lung tissue homogenates of ALI mice were determined by qRT-PCR. G Lung tissues of mice with various treatments were stained with CXCL1 antibody. Scale bars, 250 μm (PBS: n = 3, LPS: n = 4, LPS+Ctrl-shRNA MuSCs: n = 5, LPS + IL4I1-shRNA MuSCs: n = 5). Data were shown as means ± SEM. Data were representative of three experiments with similar results. For multiple group comparison, statistical analysis was performed by one-way ANOVA 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/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-4I1 by Western Blot IFN-gamma and TNF-alpha increase the expression of IL4I1 in MuSCs through NF-kappa B and STAT6 pathway.A Volcano plot of differentially expressed genes in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. B Volcano plot of differentially expressed genes in MSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h using RNA-seq. C The mRNA expression of IL4I1 in MSCs and MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was assayed by qRT-PCR. D The concentration of IL4I1 in the supernatants of MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was measured by ELISA. E The protein expression of IL4I1 and beta -ACTIN (loading control) in MuSCs after the stimulation of IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h was determined by western blotting. F The mRNA and protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of AS1517499 were respectively assayed by qRT-PCR and western blotting. G The protein expression levels of IL4I1 in MuSCs stimulated with IFN-gamma and TNF-alpha (10 ng/ml each) for 24 h in the presence or absence of BAY117082 were assayed by western blotting. Data were shown as means ± SEM. Data were representative of three experiments with similar results. For two-group comparison, statistical analysis was performed by Student’s t test. ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37507432), licensed under a CC-BY license. Not internally tested by R&D Systems.