Figure 4.

c-Yes-silencing and 5FU inhibits YAP nuclear translocation. A- Western blot analysis of c-Yes, YAP and β-catenin in c-Yes silenced Sh1 and Sh2 cells as compared to control Scr cells. B- Percentage of cells arrested at the quiescent G0 state in c-Yes silenced Sh1 and Sh2 cells. Results are expressed in percentage of quiescent cells in G0 state. C- Immunoblot of YAP and transcription factor Sp1 (nuclear marker) in nuclear and cytoplasmic fractions from c-Yes silenced Sh1 cells and control Scr cells. D- Immunoblots of YAP and β-catenin in control and 5FU-treated (2×IC₅₀, 4 days) 5F31 cells. E- Impact of subsequent 5FU treatement (2×IC₅₀, 4 days) on the nuclear distribution of YAP in 5F31 cells. Western blots are representative of at least two independent experiments. F- Subcellular localization of c-Yes by confocal microscopy analysis in control and 5FU-treated 5F31 cells. G- Coimmunoprecipitation (IP) of c-Yes with the anti-YAP antibody followed by immunoblotting using the c-Yes antibody and the YAP antibody in control and 5FU-treated 5F31 cells. Non-specific c-Yes immunoprecipitation was monitored using a non-relevant IgG.