Extended Data Figure 9: Role of Yap- and Egfr-dependent signalling in Apc mutant cells.
From: Yap-dependent reprogramming of Lgr5+ stem cells drives intestinal regeneration and cancer
a, qPCR analysis of selected Yap regulated genes comparing fold change between Apc mutant Yap+/Δ and YapΔ/Δ organoids. Error bars indicate s.e.m.; n > 4 (n represents the number of independent organoid cultures per genotype per mouse analysed for each gene). b, Western blot analysis showing expression of pErk1/2 in Yap+/Δ;ApcΔ/Δ (het), YapΔ/Δ;ApcΔ/Δ (KO) and doxycycline-treated YapTg;ApcΔ/Δ organoids (Tg). Effects of PD153035 and U0126 treatment on pErk1/2 levels are also shown (n = 3; n represents the number of separate organoid cultures analysed per genotype). c, d, Yap+/Δ;ApcΔ/Δ and YapΔ/Δ;ApcΔ/Δ mice were injected with tamoxifen and crypts harvested 48 h later. Organoids were grown in medium lacking Rsp1, Noggin and Egf and supplemented with 0.5 μg ml−1 of recombinant Ereg. Immunofluorescence shows that Ereg treatment strongly enhances Edu incorporation in both control and YapΔ/Δ;ApcΔ/Δ (middle panels). Yap stainings (right panels) show that Ereg-stimulated YapΔ/Δ;ApcΔ/Δ organoids lack endogenous Yap. Images are representative of at least three cultures derived from individual mice. Scale bars, 70 μm.
