Fig. 4

Organoid formation in the presence of Wnt regulators. a Cells isolated from β1-integrin-flox mice treated without or with 4-OHT were incubated with either 100 ng/ml Wnt3a or 50 nM glycogen synthase kinase 3 inhibitor (GSK3i), left for 10 days in organoid media to form solid and hollow organoids, and the percentage of solid organoid-forming cells was determined. Error bars represent SEM (n = 2). ns Non-significant. b Quantification of the hollow organoids that formed in the experiment shown in (a). c Role of integrin-Rac signalling in the formation of solid MEC organoids, after treating control, integrin-depleted cells or those also expressing active-Rac, and treated with Wnt3a or GSK3i (n = 3). Error bars = SEM (statistical significance determined by one-way analysis of variance). d Role of integrin-Rac signalling in the formation of hollow MEC organoids. e Inhibiting Rac1 with EHT1864 prevents solid organoid formation. f Role of EHT in hollow organoid formation. * = p < 0.05, ** = p < 0.01, *** = p < 0.001