Mouse Oncostatin M/OSM Antibody

Oncostatin M/OSM in Mouse Embryo. Oncostatin M/OSM was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c, section through spinal cord) using Mouse Oncostatin M/OSM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-495-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Oncostatin M/OSM in Mouse Embryo. Oncostatin M/OSM was detected in immersion fixed frozen sections of mouse embryo using Mouse Oncostatin M/OSM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-495-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Cell Proliferation Induced by Oncostatin M/OSM and Neutralization by Mouse Oncostatin M/OSM Antibody. Recombinant Mouse Oncostatin M/OSM (Catalog # 495-MO) stimulates proliferation in the NIH‑3T3 mouse embryonic fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse Oncostatin M/OSM (15 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Oncostatin M/OSM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-495-NA). The ND₅₀ is typically 0.6-3.0 µg/mL.

Detection of Mouse Oncostatin M/OSM by Western Blot OSM suppresses SMAD signaling. a After pretreatment with the OSM (10 ng per ml) or IL6 (20 ng per ml), C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression level of alpha SMA mRNA was calculated. The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (2, 6) = 97.35). b Immunoblot analysis of phospho-ERK1/2, total ERK1/2, phospho-SMAD2 linker (Ser245/250/255) and total SMAD2 protein. Thirty minutes after U0126 (20 nM) pretreatment, C3H/10T1/2 cells were stimulated with OSM (10 ng per ml) or IL6 (20 ng per ml) for 30 min. and collected for the analysis. U0126: a selective MEK1/2 inhibitor. c Immunoblot analysis of phospho-SMAD2 C-terminal (Ser465/467) and Lamin A/C. Thirty minutes after OSM (10 ng per ml) pretreatment, C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and collected at indicated time points. d After pretreatment with U0126 (20 μM, 60 min) and OSM (10 ng per ml, 30 min), C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression level of alpha SMA mRNA was calculated. Two-tailed t-test with Welch’s correction was used for the statistical analysis (t = 3.212, df = 3.172). Data show the mean and the standard deviation (error bar) of technical triplicates from a representative experiment (a, d). *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31249305), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Oncostatin M/OSM by Immunohistochemistry Oncostatin M from hypoxic MΦ inhibits fibroblast activation. a Supernatants were collected from thioglycollate-elicited peritoneal macrophages (TEPMs) under normoxic or hypoxic condition. After pretreatment with the supernatants, C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated (right). The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (2, 6) = 6.019). b We performed transcriptome analysis in isolated primary TEPMs from hematopoietic/endothelial-specific HIF-1 alpha conditional knockout mice (HIF-1 alpha flox/flox; Tie2-cre+/− mice; HIF-1 alpha KO) or cre negative littermate control. Flow chart shows the strategy to identify hypoxia-inducible secretory factors (left). Table illustrates the identified 11 hypoxia-inducible secretory factors (right). c The effect of each hypoxic inducible secretory factor in fibroblasts activation was examined (20 ng per ml for CTGF, CXCL2, CXCL3, LIF and 10 ng per ml for OSM). Thirty minutes after pretreatment with hypoxia-inducible secretory factors, C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated. The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (5, 12) = 68.07). d The effect of oncostatin M (OSM) (10 ng per ml) in primary cardiac fibroblasts activation was examined. Thirty minutes after pretreatment with OSM (10 ng per ml, 12 h), C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated. Two-tailed t-test with Welch’s correction was used for the statistical analysis (t = 10.89, df = 2.208). Data show the mean and the standard deviation (error bar) of technical triplicates from a representative experiment. e Immunohistological staining were performed using cardiac tissues from TAC operated mice (day 3). OSM (left), CD11b (right). Scale bar = 50 μm. *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31249305), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Oncostatin M/OSM by Immunohistochemistry Oncostatin M from hypoxic MΦ inhibits fibroblast activation. a Supernatants were collected from thioglycollate-elicited peritoneal macrophages (TEPMs) under normoxic or hypoxic condition. After pretreatment with the supernatants, C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated (right). The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (2, 6) = 6.019). b We performed transcriptome analysis in isolated primary TEPMs from hematopoietic/endothelial-specific HIF-1 alpha conditional knockout mice (HIF-1 alpha flox/flox; Tie2-cre+/− mice; HIF-1 alpha KO) or cre negative littermate control. Flow chart shows the strategy to identify hypoxia-inducible secretory factors (left). Table illustrates the identified 11 hypoxia-inducible secretory factors (right). c The effect of each hypoxic inducible secretory factor in fibroblasts activation was examined (20 ng per ml for CTGF, CXCL2, CXCL3, LIF and 10 ng per ml for OSM). Thirty minutes after pretreatment with hypoxia-inducible secretory factors, C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated. The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (5, 12) = 68.07). d The effect of oncostatin M (OSM) (10 ng per ml) in primary cardiac fibroblasts activation was examined. Thirty minutes after pretreatment with OSM (10 ng per ml, 12 h), C3H/10T1/2 cells were stimulated with TGF-beta 1 (2.5 ng per ml, 12 h) and the relative expression of alpha SMA mRNA was calculated. Two-tailed t-test with Welch’s correction was used for the statistical analysis (t = 10.89, df = 2.208). Data show the mean and the standard deviation (error bar) of technical triplicates from a representative experiment. e Immunohistological staining were performed using cardiac tissues from TAC operated mice (day 3). OSM (left), CD11b (right). Scale bar = 50 μm. *p < 0.05 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31249305), licensed under a CC-BY license. Not internally tested by R&D Systems.