Paxillin depletion promotes MT deacetylation through activation of HDAC6. (A) Western blots of MDA-MB-231 cells treated with control or paxillin RNAi as indicated, with 2 µM tubacin for 4 h or HDAC6 RNAi. (B) RNAi of SIRT2, another α-tubulin deacetylase, had minimal effect on global α-tubulin acetylation as assessed by Western blot analysis. (C–E) Western blots (C), quantitation (D), and images (E) of α-tubulin deacetylation after tubacin washout. Boxed regions indicate the area used for the respective zoomed images. LUT, lookup table. (F and G) Western blots (F) and quantitation (G) of the time course of α-tubulin acetylation after the addition of 2 µM tubacin. Data are represented as the mean ± SEM. *, P < 0.05 from n = 3 individual experiments.