Paxillin regulates MT acetylation in malignant and normal cell types. (A–C) Western blot (A) quantitation (B), and images (C), indicating a reduction in α-tubulin acetylation upon paxillin depletion in MDA-MB-231 cells. Boxed regions indicate the area of the merged image used for the zoom. RNAi-mediated Hic-5 protein depletion did not significantly affect α-tubulin acetylation (P = 0.65). n = 3 individual experiments. (D) Images of microtubule (MT) acetylation in Hs578T and HFF cells after paxillin depletion. (E and F) Images (E) and quantitation (F) of mean fluorescence intensity (MFI) of acetylated α-tubulin immunofluorescence in paxillin^−/− MEFs reexpressing GFP-paxillin, as indicated by the asterisks. Pseudocolored lookup table (LUT) images highlight changes in acetylated α-tubulin pixel MFI. n = 3 individual experiments, and MFI from a minimum of 21 cells was quantified. Data are represented as the mean ± SEM. *, P < 0.05; ***, P < 0.0005.