Fig. 5: IEC-specific overexpression of constitutively activated STAT6 (STAT6vt) promotes tuft and goblet cell differentiation.

2-month-old male WT and TgSTAT6vt mice (including line-10, line-13 and line-18) were subjected to the following assays. a The strategy for creating TgSTAT6vt mice. b, c Western blot (b) and qPCR analyses (c) of TgSTAT6vt expression in the jejunal IECs. n = 7 mice/group in (c); p = 0.0013 (line-10), <0.0001 (line-13), <0.0001 (line-18). d Representative H&E images of jejunum tissues. e Analyses of jejunal villus length and crypt depth. n = 30 villi or crypts/group; p < 0.0001 (villus length, crypt depth). f qPCR analysis of IEC markers expression in the jejunal IECs. Dclk1, Lyz1, n = 4 mice/group; Trpm5, Retnlb, n = 5 mice/group; Slc5a1, n = 5 (WT) and 4 (Tg) mice; Chga, Lgr5, n = 4 (WT) and 3 (Tg) mice; p = 0.0259 (Dclk1), 0.0044 (Trpm5), 0.0491 (Retnlb), 0.0322 (Lyz1). g Tuft and goblet cells were examined by DCLK1 immunostaining and Alcian blue staining, respectively in the jejunum (200X). (h) Quantification of tuft and goblet cells shown in (g). n = 4 mice/group; 20 crypt-villus units counted for each mouse; p = 0.0018 (tuft cell) and 0.0126 (goblet cell). i Tuft and goblet cells were labeled by anti-DCLK1 and anti-MUC2, respectively, in frozen sections of organoids (200X). j Quantification of tuft and goblet cells shown in (i). n = 10 organoids/group; p < 0.0001 (tuft cell, goblet cell). Data are presented as mean ± SEM. All p values were generated by two-tailed unpaired t test. *p < 0.05. Scale bars, 100 µm in (d, g); 50 µm in (i). Source data are provided as a Source Data file.