Fig. 3.

GAK is required for the generation of chromatin-derived microtubules. a–c U2OS cells were transfected with indicated siRNAs. Twenty-four hours after transfection, cells were re-transfected, and 48 h after the first transfection, cells were treated with nocodazole for 6 h, washed four times with medium to remove nocodazole, and fixed after 3 min (a, upper panels and b, c) or after 10 min (a, lower panels) and stained for α-tubulin and γ-tubulin. DAPI was used to visualize the DNA. a Representative images. b, c Quantifications. d Cells were transfected as in (a–c) but were fixed after 48 h without nocodazole treatment. Cells were then stained for HURP and α-tubulin, and DNA was stained with DAPI. HURP localizes specifically to DNA-proximal microtubules both in control cells and in GAK-depleted cells. Graphs in (b, c) show averages of three independent experiments with 25 cells scored per condition per experiment. Error bars represent standard deviations. Scale bars in (a, d) indicate 5 µm