Fig. 4

Loss of sumoylation results in severe mitotic dysfunction with significant DNA damage upon mitosis entry. A Jeko cells or Z-138 cells (bottom) were transduced with a H2B-GFP construct. Cells were synchronized with palbociclib (500 nM, 24 h), washed and released into DMSO or TAK-981 (100 nM). GFP and phase-contrast images were acquired every 5 min over 24 h. (Top) Representative series of images of Jeko cells transitioning through mitosis in the presence of TAK-981. Jeko (bottom left) or Z-138 (bottom right) were followed and the result of mitosis for individual cells was quantified (n = 50 per group, two independent experiments for each). B Representative confocal microscopy images of mitotic Jeko cells (top) for alpha tubulin (green), gamma-tubulin (red), and DAPI (blue) and mitotic Jeko (bottom left, middle) and Z-138 cells (bottom right) for alpha tubulin (green), CENP-A (red), and DAPI (blue). C, D Jeko (left) and Z-138 cells (right) were synchronized with palbociclib (500 nM) and treated with either DMSO or TAK-981 (100 nM). Lysates were prepared at the indicated time points after washout. The corresponding phase of the cell cycle based on the DNA profiles obtained from fixed, PI stained cells from each of the time points is shown (see Additional file 1: Figures S7, S8). Lysates were blotted for the indicated proteins