Fig. 4: AR activates MLCK signaling pathway and alters intestinal epithelial barrier function in naïve C57BL/6 mice and murine intestinal organoids.
a Mlck mRNA levels. b Representative western blot analysis of pMLCSer19, MLC, and β-actin. Uncropped blots are provided in the Source Data. c Relative densitometry (pMLC/MLC) (n = 3 mice/group). d Representative bright field (BF) images of mouse colonic organoids treated for 24 h with or without AR (1 μmol L−1) following pre-treatment for 1 h with or without TNF-α (10 ng/mL). e Percentage of disrupted organoids. f Mlck mRNA levels in 2D monolayer derived from murine colonic organoids. g Representative western blot analysis of ZO-1 and β-actin. Uncropped blots are provided in the Source Data. h Relative densitometry (ZO-1/β-actin) (n = 3 per group). i Tjp1 mRNA levels. j Representative images of PAS–stained colon sections; scale bar: 50 µm. k Number of PAS+ colonic goblet cells per 10 crypts. l Muc2 mRNA levels. a–l (except e and f) Data were analyzed by two-tailed unpaired Student’s t-test and are expressed as mean or mean ± SD (n = 4 for Chow; n = 5 for AR). e, f Data were analyzed by one-way ANOVA with post hoc Bonferroni’s test. Data are expressed as mean ± SD and representative of 2 independent experiments. Significance denoted by *p < 0.05, #p < 0.05 unless otherwise provided, where *p < 0.05 versus untreated 2D monolayer derived from murine colonic organoids, and #p < 0.05 versus TNF-α treated 2D monolayer derived from murine colonic organoids. Source data are provided as a Source Data File.
