Fig. 4: Induction of OSMR in epithelial cells is dependent on STAT3.
From: The IL-22–oncostatin M axis promotes intestinal inflammation and tumorigenesis
a, H. hepaticus + anti-IL-10R colitis was induced in wild-type mice for 14 days. Protein analysis of IECs from steady-state (n = 4) and H. hepaticus + anti-IL-10R (n = 5) colitic mice was conducted. The western blot depicts phosphorylated STAT1 (pSTAT1), STAT1, pSTAT3, STAT3 and β-actin for representative samples from two experiments. b,c, Immunofluorescence staining for pSTAT1 and pSTAT3 in steady-state (n = 7) and inflamed (n = 10) mouse samples (b) with subsequent quantification (c) in the epithelium; scale bar, 20 μm. Hpf, high-power field. Data were pooled from two experiments. d,e, H. hepaticus + anti-IL-10R colitis was induced in wild-type mice for 7 days, and mice were treated with anti-IL-22 (clone 8e11, n = 4) or isotype control (mouse IgG1, GP120:9709, n = 5) on days 0 and 3 of colitis induction. Protein analysis of IECs from steady-state and H. hepaticus + anti-IL-10R colitic mice and anti-IL-22 or isotype-treated inflamed mice was performed. d, Western blot depicting STAT3 phosphorylation in epithelial cells from anti-IL-22- or isotype-treated colitic mice. Data are representative of two independent experiments. e, Quantification of STAT3 phosphorylation using ImageJ. The relative band intensity of blots from inflamed mice was normalized to that of steady-state untreated mice. f,g, Colitis was induced in VilcreERT2Stat3fl/fl (IECΔStat3) or VilcreERT2Stat3fl/wt (IECWT; f) and Stat1–/– or Stat1+/+ (g) mice using the H. hepaticus + anti-IL-10R model for 7 days. qPCR analysis was performed to assess the expression of Osmr and Reg3g in epithelial cells from inflamed mice normalized to epithelial cells isolated from untreated respective control mice. Data are representative of one experiment; n = 5–7 mice per genotype. h, Mouse colon organoids generated from Vilcre+Stat3fl/fl (IECΔStat3) or Vilcre-Stat3fl/fl (IECWT) mice and stimulated with 10 ng ml–1 IL-22. Relative expression of Osmr was analyzed by qPCR. Data are representative of three independent experiments from three independent biological replicates. P values (two-tailed) were calculated using a Mann–Whitney test for c and e–h.
