Fig. 3

RhoGAPs depletion induces junction recovery. a Staining for ZO-1, E-cadherin and DNA in HT29 cells transfected with CTRL si or a mix of GRAF2 si #2, p190A si #2 and p190B si #2 (GAPs si #2). Relative ZO-1 positive areas are also shown. n = 77, 72, and 69 fields for cells transfected with CTRL si, GAPs si #1 (a mix of GRAF2 si #1, p190A si #1 and p190B si #1) and GAPs si #2, respectively, pooled from two independent experiments. ****P < 0.0001 by two-tailed Mann–Whitney U-test. b Staining for ZO-1 and DNA in HT29 cells transfected with CTRL siRNA or GAPs si #2 or a mix of GAPs si #2 and GEF-H1 si #1. Relative ZO-1 positive areas are also shown. n = 45, 43, 45, and 42 fields for cells transfected with CTRL si, GAPs si #2, a mix of GAPs si #2 and GEF-H1 si #1 or a mix of GAPs si #2 and GEF-H1 si #2, respectively, pooled from two independent experiments. ****P < 0.0001 by two-tailed Mann–Whitney U-test. c Immunostaining for α-tubulin in HT29 cells transfected as in a. Images were obtained with Airyscan. d Colo205 cysts transfected with CTRL or GAPs si, which were stained for F-actin, ZO-1 and DNA. Cells were cultured in Matrigel for 3 days. The ratio of cysts with a single, multiple and no lumens is shown. n = 207 cysts for CTRL si, 178 and 181 cysts for GAPs si #1 and #2, respectively, pooled from two independent experiments. See also Supplementary Fig. 3