Figure 5From: Topotecan synergizes with CHEK1 (CHK1) inhibitor to induce apoptosis in ovarian cancer cellsCombined treatment with TPT and PF477736 showed increased apoptotic activities and DNA damage. A. Apoptotic activity was measured by Caspase3/7 activity assay. Cells were treated with TPT and/or PF477736 for 16 hours with indicated drug concentrations and Caspase3/7 activity was measured relative to untreated samples in triplicates. B. Cells were treated at the same concentrations as in Caspase3/7 assay for 16 or 24 hours and total cell lysates were prepared for Western blotting. GAPDH was used as a loading control. C. Eight hundred thousand cells were plated in 60 mm plates 8 hours prior to drug treatment. Cells were treated for 16 hours with TPT (0.2 μM) and/or PF (0.5 μM). G0/G1, S, and G2/M phases were measured based on staining of APC-BrdU and 7-AAD by flow cytometry. Cell cycle was analyzed by FlowJo software.Back to article page