Figure 2.

TNF activates YAP through Rho family of GTPases. HEK293 were cultured as described in Figure 1 . Cells were pre-treated with Y27632 at 10 µM or C3 exoenzyme cell permeable at 2 µg/ml for 2 h before adding TNF for 48 h [24 h in (C) and (G)]. (A) Representative airyscan confocal z-stack max intensity images of YAP (immunofluorescence (IF) technique, green), phalloidin (actin, red), DAPI (nucleus, blue), and merged images. (B) Corresponding IF quantification of YAP nucleo-cytoplasmic ratio. (C) and (G) Luciferase reporter assay of TEAD transcription factor activity. (D–F) RT-qPCR results for CYR61 (D), CTGF (E) and ANKRD1 (F), results were normalized to HPRT expression. (H) Representative airyscan confocal z-stack max intensity images of YAP [immunofluorescence (IF) technique, green] alone or with phalloidin (actin, red) and DAPI (nucleus, blue). For confocal images, luminosity and contrast were enhanced identically for each image for clarity purpose. Results are representative of three independent experiments with 2 to 3 biological replicates for each experiment (n = 6 to 9 per group) with T-test or one-way ANOVA test and FDR corrected for multiple comparisons post hoc tests performed between conditions: *p < 0.05; **p < 0.01; ***p < 0.001. Data are expressed as fold change vs. control and presented as individual values with mean ± SD.