Figure 7.

LPA₂ promotes the growth of Lpar5^cKO enteroids via activation of the PI3K-Akt pathway. (A) Lpar5^cKO (Lpar5^f/f;Villin-Cre) enteroids were cultured in the presence of LPA₂ inhibitor H2L5186303 (H2L5), PI3K inhibitor LY294002 (LY), or both. Representative images of enteroids taken on D0 and D2 are shown. Bar = 50 μm. (B) Growth of enteroids was quantified by determining the average surface area of enteroids. * p < 0.05, **** p < 0.0001, and ns = not significant by two-way ANOVA with Tukey’s multiple comparison test. (C) Representative immunofluorescence images for p-Akt (upper panels) and Akt (lower panels) are shown. DAPI = blue. Images are representative of three independent experiments. Bar = 50 µm. (D) Ratios of p-Akt to total Akt fluorescence intensity (mean ± SD) are shown. n = 10. **** p < 0.0001 and ns = not significant by one-way ANOVA with Tukey’s multiple comparison test. (E) Growth of Lpar2^−/− enteroids in the presence of LPA₅ inhibitor AS2717638 (AS27), PI3K inhibitor LY294002 (LY), or both were determined and average surface areas of enteroids (mean ± SD) under each condition are shown. * p < 0.05, **** p < 0.0001, and ns = not significant by two-way ANOVA with Tukey’s multiple comparison test.