Figure 4: Acetylation of microtubules decreases adhesion disassembly and promotes adhesion maturation.

(a) Representative confocal images of HFFs expressing GFP (control) or different tubulins as indicated immunostained for FN (green) and paxillin (red). Front of cell (hatched box) enlarged in the right panels. (b) Quantification of elongated adhesions in a from a minimum of 25 cells (n=3). (c) Representative confocal images of HFFs expressing the indicated tubulin mutants and immunostained with anti-α₅ integrin (left panels) or incubated overnight with Alexa Fluor 562-labelled FN (right panels; n=nucleus). (d) Maximally projected confocal images of matrices derived from cells expressing the indicated tubulin mutants and immunostained for human FN (antibody 3B8). (e) Human FN from matrices described in d quantified by western blot as relative intensity (n=4). (f) Representative confocal spinning disc images of HFFs expressing different tubulins as indicated with mApple-paxillin; see also Supplementary Movie 1. (g) Scatter plot of disassembly rate of adhesions from f. Data are mean (red bars) ±s.e.m. *P<0.05 and **P<0.01. Two-way analysis of variance (ANOVA) followed by Tukey post test was used for b, e and g. Scale bar, 20 μm (a; left panels); scale bar, 10 μm (a; right panels and c); scale bar, 100 μm (d); scale bar, 5 μm (f).