Fig. 5

Fig. 5 AMPK-dependent phosphorylation of YAP at S61 regulates autophagy upon fluid flow.

a Representative images of P-YAP (S61), YAP and Actin proteins levels in KECs subjected to flow (shear) or not (static) during 24 h, by western blot analysis. b The ratio of P-YAP (S61) to YAP was determined by densitometry, relative to panel a. Data show the mean ± s.e.m.; n = 3 independent experiments, two-sided t-test. c, d Representative images (c) and quantification (d) of LC3 puncta in KECs expressing a wild-type (WT) or a mutant (S61A) form of GFP-YAP, subjected to shear stress during 24 h in the presence or absence of chloroquine (CQ). e, f Representative images (e) and quantification (f) of YAP nuclear localization in KECs treated or not with dorsomorphin (Dorso.), subjected to flow (shear) or not (static) during 24 h. Scale bars, 10 µm. g, h Representative images (g) and quantification (h) of lipid droplets (Bodipy) in KECs, after transfection with a control siRNA (si Scr) or siRNA against ATG5, treated or not with dorsomorphin (Dorso.) and subjected to flow (shear) or not (static) during 24 h. i Representative images of P-YAP (S61), YAP, LKB1 and Actin proteins levels in KECs after transfection with a control siRNA (si Scr) or siRNA against LKB1 subjected to flow (shear) during 24 h, by western blot analysis. j The ratio of P-YAP (S61) to YAP was determined by densitometry, relative to panel i. k Representative images of P-YAP (S61), YAP, α1 AMPK, total AMPK (α1/α2) and Actin proteins levels in kidney samples from wild-type (WT) or AMPK α1 KO mice, by western blot analysis. l The ratio of P-YAP (S61) to YAP was determined by densitometry, relative to panel k. m The ratio of AMPK to Actin was determined by densitometry, relative to panel k. d, h Data show the mean ± s.e.m.; n = 24 individual data points from 3 independent experiments, two-sided t-test. Scale bars, 10 µm. f, j Data show the mean ± s.e.m.; n = 5 (f) or n = 4 (j) independent experiments, two-sided t-test. l, m Data show the mean ± s.e.m.; n = 3 kidney samples from WT or AMPK α1 KO mice, two-sided t-test. Source data are provided as a Source Data file.