MT detyrosination does not interfere with global kMT dynamics . (A) Representative images of DIC and time-lapse fluorescent images of metaphase spindles in control, TTL-deficient (siTTL) and MCAK-deficient (siMCAK) U2OS cells stably expressing photoactivatable-GFP/mCherry-tubulin before photoactivation (Pre-PA) and at the indicated time points (min) after photoactivation of GFP-tubulin fluorescence. Scale bars, 5 µm. (B) Normalized fluorescence intensity after photoactivation in all conditions for metaphase cells. Data points represent mean ± SD. (C) Relative distribution of kMTs and non-kMTs obtained for each experimental condition after photoactivation (D) Calculated kMT t_1/2 under different conditions. Bars indicate mean values/cell, and error bars represent SD. Control-prometaphase: n = 31, pool of five independent experiments; control-metaphase: n = 32 cells, pool of six independent experiments; siTTL-prometaphase: n = 28, pool of six independent experiments; siTTL-metaphase: n = 17 cells, pool of three independent experiments; siMCAK-prometaphase: n = 15, pool of three independent experiments; siMCAK-metaphase: n = 30 cells, pool of two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, nonsignificant; unpaired two-tailed t test.