Fig. 5.

Microtubule acetylation regulates TGF-β1-induced nuclear translocation of YAP, resulting in the promotion of Smad2 transcriptional activation. a Subcellular fractionation in WT and α-TAT1 KO MEFs incubated on soft and stiff matrices and stimulated with TGF-β1. Each fraction was used for western blotting with antibodies specific for phospho-Smad2/3 and YAP. GAPDH and Lamin A were used as markers for cytosolic and nucleic fraction, respectively. b, c Luciferase reporter assay for transcriptional activity of Smad (b; SBE2-Luc) and YAP (c; 8xGTIIC-Luc) in YAP and Smad2 KD cells incubated on soft or stiff matrices. Data are represented as mean of ± S.D. from three independent experiments (n = 6). SBE2-Luc: one-way ANOVA, F_{5, 30} = 35.25 (soft), F_{5, 30} = 98.93 (stiff). 8xGTIIC-Luc: one-way ANOVA, F_{5, 30} = 59.00 (soft), F_{5, 30} = 85.86 (stiff). d WT and α-TAT1 KO MEFs were transfected with the indicated plasmids (GFP or YAP(5SA)-GFP) for 24 h. WT and α-TAT1 KO MEFs were seeded on fibronectin-coated soft matrix and incubated for 8 h. Graphs show the relative luciferase activity of YAP and Smad. Data represent the mean of three independent experiments ± S.D. (n = 6). 8xGTIIC-Luc: one-way ANOVA, F_{3, 20} = 311.6, SBE2-Luc: one-way ANOVA, F_{3, 20} = 95.15. ***p < 0.005, n.s. not significant