Fig. 3

Loss of sumoylation results in mitotic dysregulation with accumulation of polyploid cells A (left). Volcano plot of genes with a statistically significant association with SAE1 and SAE2 (FDR < 0.01). (right) DAVID analysis was performed on the statistically significant gene list. The top gene groups and enrichment scores are shown. B (right). Normal and activated B-cells and MCL cell lines were treated with DMSO, 50 nM or 100 nM of TAK-981 and cells were collected daily. Cells were fixed, stained with PI and cell cycle analysis was performed. Representative DNA profiles are shown for the indicated cell lines and primary samples treated with 100 nM of TAK-981. Quantification of 2n cells relative to DMSO control (top, right) at 24 h and percentage polyploid cells (bottom, right) after 48 h of TAK-981 treatment at the indicated concentrations (n = 3 independent experiments per cell line). C Primary MCL samples were treated and analyzed as in B (n = 3 independent experiments per sample). D (Left) The indicated cell lines were treated with palbociclib (200 nM) for 24 h and then released into DMSO, palbociclib, TAK-981 or palbociclib + TAK-981. (right top) Percentage viable cells was quantified after 72 h from the time of release from the initial palbociclib treatment (n = 3 independent experiments for each cell line, * p < 0.05). (right bottom) Western blot confirming equivalent losses of sumoylated proteins with TAK-981 in the presence or absence of TAK-981