Fig. 4.

Wee1 inhibition induces Cdk1-dependent cytokinesis failure. (A) FUCCI-MDA-MB-231 cells were treated and imaged as in Fig. 3 and Fig. S6A. The percentages of multinucleated MDA-MB-231 and SKOV3 cells were quantified at indicated time points. (B) Representative immunofluorescence images of untreated MDA-MB-231 colonies or colonies that survived combined MK-1775 (500 nM) and SU-9516 (1 μM) treatment. Cells were fixed at 14 d after treatment and stained for CD44 (red) and DNA (white). (C) pLKO.puro transduced MDA-MB-231 and SKOV3 cells were treated with MK-1775 (500 nM) for 14 d and stained as in B. Percentages of multinucleated cells were quantified and presented. (D) MDA-MB-231 cells transfected with CDK1-AF or CDK1-WT were treated with MK-1775 (4 μM) and subsequently imaged every 7 min for 62 h. Mitotic exit was analyzed for >15 CFP-positive cells. (E) A model for Wee1 inhibition causing cytotoxicity and genomic instability.