Figure 7.

Transforming growth factor-β (TGFβ)/bone morphogenetic protein (BMP) signaling via SMAD4 modulates the expression of Cldn genes by disparate mechanisms. A: wild-type colonoids treated with TGFβ1/BMP2 (gray bars/open boxes) or vehicle (white bars/closed circles) for the indicated periods of time. B: wild-type colonoids treated with TGFβ1/BMP2 (T/B) vs. vehicle (white bars, left), T/B vs. vehicle in the presence of actinomycin D (ActD) (light gray bars, middle), or T/B vs. vehicle in the presence of cyclohexamide (CHX) (dark gray bars, right) for 6.5 h. Fold change indicates the relative level of mRNA for indicated genes compared with vehicle-treated control. Each data point represents a single biological replicate, with biological replicates treated and harvested on different days. Experimental arms were compared using nonparametric Kruskal–Wallis test with post hoc Welch’s t test. Smad7 is used as a positive control to confirm ligand activity. Statistical significance is designated as *P < 0.05, **P < 0.01.