Fig. 5

IFN-γ inhibits Stat3 activation through Stat1 to block the differentiation of primary ECs. (a) Representative IF images of Ace. Tub staining and quantification of multiciliated ECs (n = 3). Scale bars, 50 μm. Unpaired T tests were applied. * P < 0.05. (b) Representative IF images and quantification of Foxj1-positive ECs (n = 3). Scale bars, 100 μm. Unpaired T tests were applied. ** P < 0.01. (c) Representative immunoblot images and quantification of Foxj1 protein levels on D1, 3, and 7 of EC differentiation (n = 3). Two-way ANOVA tests were applied. ** P < 0.01 and *** P < 0.001. (d) qPCR analysis of foxj1 mRNA levels on D1, 3, and 7 of EC differentiation (n = 6). Two-way ANOVA tests were applied. *** P < 0.001 and **** P < 0.0001. (e) Representative immunoblot images and quantification of Stat1 protein expression and phosphorylation activation on D1 and 3 of EC differentiation (n = 3). Two-way ANOVA tests were applied. * P < 0.05 and *** P < 0.001. (f) Representative immunoblot images of pStat1, Stat1, and Foxj1 protein levels in ECs, with representative images and statistical results for Foxj1 (n = 3). One-way ANOVA tests were applied. * P < 0.05. (g) Representative IF images of Ace. Tub staining and quantification of multiciliated ECs (n = 3). Scale bars, 100 μm. One-way ANOVA tests were applied. ** P < 0.01 and *** P < 0.001. (h) Representative immunoblot images of Stat1 protein levels and phosphorylation activation in HEK293T cells (n = 3). (i) Quantification of the luciferase activity in HEK293T cells (n = 3). Two-way ANOVA tests were applied. (j) Representative immunoblot images and quantification of pStat3 and Stat3 expression levels in ECs (n = 3). Unpaired T tests were applied. * P < 0.05. (k) Representative immunoblot images and quantification of pStat3 and Foxj1 protein expression levels in ECs (n = 3). One-way ANOVA tests were applied. * P < 0.05 and *** P < 0.001. (l) Representative IF images of Ace. Tub staining and quantification of multiciliated ECs. Scale bars, 100 μm. (n = 3). One-way ANOVA tests were applied. *** P < 0.001