Mouse Oncostatin M/OSM Antibody

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.

Detection of Mouse Oncostatin M/OSM by Flow Cytometry

UMAP visualization of skin scRNAseq analysis. (A) UMAPs of scRNAseq from skin punches of FT+/− and WT neonates treated with Alt or Alt/PNE with cluster-specific cell type annotation. (B) UMAP projection of skin cells expressing Osm (green), Areg (blue), and Il33 (red) above the threshold expression value (Seurat log Normalized) >1.5. Annotated cell type designation to numbered clusters in (A) is listed as follows; Dendritic, dendritic cells; Endo, endothelial cells; Fib, fibroblasts; ILC, innate lymphoid cells; KC, keratinocytes; Mac, macrophages; PMN, neutrophils; SM, smooth muscle cells; Stem, stem cells. (C) Cell clusters with abundant gene expression of Osm, Areg, and Il33. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35387035), licensed under a CC-BY license. Not internally tested by R&D Systems.