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. 2016 Oct 18;6:35624. doi: 10.1038/srep35624

Figure 3. Stochastic acetylation of microtubules assembled in vitro.

Figure 3

(A) MTs assembled in vitro from HeLa cell-purified tubulin dimers were incubated in the presence of 4 μM recombinant αTAT1 for the indicated time period before being fixed and stained for total tubulin (red) and acetylated K40 tubulin (green). Scale bar: 5 μm. (B) Average fluorescence intensity distribution of total tubulin staining (red) and acetylated K40 tubulin (green) along MTs (MTs tip at x = 1 μm) after a 2 min incubation period with recombinant αTAT1 as in A. Data are represented as mean ± S.E.M. (C) Average fluorescence intensity distribution of acetylated K40 tubulin along MTs (MTs tip at x = 1 μm) after a 0 (black), 2 (red) or 4 (green) min incubation period with recombinant αTAT1 as in A. Data are represented as mean. The S.E.M was omitted for clarity. (E,D) Theoretical average acetylation marks distribution along MTs (MTs tip at x = 1 μm) after the indicated incubation period with 4 μM αTAT1 without (E) or with (D) allowed lateral access. Data were convoluted with a 250 nm width Gaussian point-spread-function (PSF) to better qualitatively compare with the microscopy results. The color-coded legend indicates the time in minutes since entry into the unacetylated lumen begins.