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. 2014 Sep 25;5(21):10546–10557. doi: 10.18632/oncotarget.2508

Figure 6. CHK1i cooperates with WEE1i in promoting mitotic catastrophe.

Figure 6

(A) Combining CHK1i and WEE1i induces extensive cell cycle disruption. HeLa cells were exposed to the indicated concentrations of CHK1i and WEE1i individually or in combination. After 24 h, the cells were harvested and analyzed with flow cytometry. (B) Combining CHK1i and WEE1i induces mitotic catastrophe. Cells were treated as described in panel (A). Lysates were prepared and analyzed with immunoblotting. Uniform loading of lysates was confirmed by immunoblotting for actin. The cells were also harvested for trypan blue exclusion assay (bottom panel, average ±SD of triplicated counting). Combination of CHK1i and WEE1i reduced viability (** P < 0.01; * P < 0.01; Student's t-test). (C) Co-inhibition of CHK1 and WEE1 promotes extensive mitotic delay and cell death. HeLa cells expressing histone H2B-GFP were incubated with CHK1i (100 nM) or WEE1i (100 nM) individually or in combination. Individual cells were then tracked for 24 h with time-lapse microscopy. Each horizontal bar represents one cell (n = 50). Grey: interphase; black: mitosis (from DNA condensation to anaphase); truncated bars: cell death. The mitotic duration was quantified (mean ±90% CI) (*** P < 0.001; Student's t-test).