Fig. 7. Disruption of epithelial integrity does not perturb WNT signaling activation.

a Cultures of WT, CTRLsh, GPC4sh5-c10 and GPC4sh2-c3 029 hiPSCs were stimulated for 6 h with 50 ng/mL of WNT3A, and analyzed for LEF1 (magenta) and ZO1 (green) proteins. Dashed areas highlight the disturbed TJs, n = 3. Scale bar: 100 μm. b Percentages of LEF1 expressing cells were quantified from staining in a. Box plots represent the median with min and max values, n = 3. c Percentages of LEF1 expressing cells as a function of the distance from zones with disrupted TJs quantified in GPC4sh5-c10 and GPC4sh2-c3 029 hiPSCs from staining shown in a. 0 μm indicates cells located within the zones of TJs disruption, +30 μm, +60 μm, +90 μm and <+90 μm indicate cells located at 0–30 µm, 30–60 µm, 60–90 µm or >90 µm from the zone of TJs disruption respectively. Data are represented as mean ± SD, n = 3. d Cultures of CTRLsh and GPC4sh5-c10 029 hiPSCs were stimulated for 6 h with 5, 10, 25 and 50 ng/mL of WNT3A, and then analyzed for LEF1 (magenta) and ZO1 (green) proteins. Outlined areas highlight the disturbed TJs, scale bar: 100 μm, n = 3. e Percentages of LEF1 expressing cells in CTRLsh and GPC4sh5-c10 029 hiPSCs from staining shown in d. Box plots represent the mean with min and max values, n = 3. f Analysis of LEF1 normalized intensity in organized and disrupted areas of GPC4sh5-c10 029 hiPSCs from staining shown in d, represented as a box and whisker plot; whiskers represent 1st and 99th percentiles, n = 3 with 7957 to 26429 single nucleus values per condition. Statistical analysis for the overall figure: (b) two-way ANOVA followed by Dunnett’s multiple comparison test, (c) linear regression analysis, (e) two-way ANOVA followed by Sidak’s multiple comparison test. P-values: (***) <0.001, (**) <0.01, (*) <0.05, ns not significant. For all panels “n” corresponds to the number of biological replicates unless stated otherwise. Source data are provided as a Source Data file.