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. 2015 Apr 19;6(15):13072–13087. doi: 10.18632/oncotarget.3865

Figure 2. Inhibition of Wee1 decreases the phosphorylation of Chk1 in gemcitabine-treated cells.

Figure 2

A, B. Panc1 and U2OS cells were treated with 1μM Wee1i or DMSO, with or without 300nM gemcitabine, for 24 h. Blots of cell lysates were stained for phosphorylation of the ATR-substrate Chk1. HSC 70 or β-Actin was stained as loading controls. C. Cells were depleted of Wee1 by transfection with 10nM siRNA for 48h, followed by gemcitabine treatment for 24 h and immunoblot analysis as in (A, B). Scrambled siRNA was used as control. D. Cells were transfected with siRNA against p53 and scrambled siRNA was used as control. After 48 h (for each condition), cells were exposed to Wee1 inhibitor in the presence or absence of gemcitabine. 24 h later, cells were harvested and immunoblotting was performed. β-Actin was stained as loading control. E, F. Cells were treated with Wee1i or DMSO, with or without gemcitabine, in the presence or absence of the pan-caspase inhibitor Z-VAD.fmk at the indicated concentrations. After 24 h, the cells were subjected to immunoblot analysis.