Fig. 8: Nocodazole treatment restores nuclear envelope morphology and diminishes DNA damage.
From: Dynamins maintain nuclear envelope homeostasis and genome stability
a Western blot of acetylated α-tubulin (top; residue Lys40), and respective quantification. Normalized to α-Tubulin (bottom). Vinculin was used as a loading control. Seven biological samples/condition. b SiR-tubulin and SPY555-DNATM stained control and dynamin TKO live cells. Scale bar 20 μm; insets 14 μm. Three independent biological replicates. Representative images of lamin A/C and γ-H2AX immunostained nuclei in control and dynamin TKO cells treated with DMSO (control) or nocodazole (c), and respective quantification of nuclear circularity (d), nuclear area (e) and γ-H2AX signal (f). Scale bar 10 μm. Four independent biological replicates; >180 cells/condition. Representative images of control and dynamin TKO cells treated with nocodazole analysed by alkaline comet assay (g), and respective quantification of DNA damage (h; Olive tail moment = [tail mean-head mean] × % of DNA in the tail). Scale bar 50 µm; insets 90 µm. Three independent biological replicates, >164 cells/condition. Data are presented as bar charts with mean ± SEM, and as box plots showing the 25th percentile (Q1), median, 75th percentile (Q3), and whiskers extending to the minimum and maximum values; statistical significance is indicated as **p < 0.01; ***p < 0.001. Statistical analyses were performed using two-tailed Student t test with Welch’s correction (a: p = 0.0003), one-way ANOVA Kruskal–Wallis test followed by post-hoc Dunn’s multiple comparisons test (d: p < 0.0001, p < 0.0001, e: p < 0.0001, p < 0.0001, f: p < 0.0001, p < 0.0001, h: p = 0.0019, p = 0.0002). Ac Acetylated, a.u. Arbitrary units, Ctrl Control, NOC Nocodazole, TKO Triple knockout.
