Combined 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.