Fig. 5

Effect of VERU-111 on cell cycle distribution and apoptosis in PanCa cells. (A) Effect of VERU-111 on cell cycle distribution of Panc-1 and AsPC-1 cells. Briefly, Cells were treated with VERU-111 for 24 h and analyzed by flow cytometric analysis using Propidium Iodide. Bar graph showing cell cycle distribution at different phases in G0-G1, S, and G2M as analyzed by MOTFIT Analysis Software (i-ii). (B) Effect of VERU-111 on protein levels of cell cycle regulatory proteins (Cyclin B1, Cdc25c, Cdc2, and pCdc2Tyr15) in Panc-1(i) and AsPC-1(ii) cells as determined by Western blot analysis. (C) Effect of VERU-111 on apoptosis induction. Briefly, cells were treated with indicated concentrations of VERU-111 for 24 h and apoptosis induction was analyzed by flow cytometry using Annexin V-7AAD Apoptosis kit. Bar graphs showing dose-dependent increase of apoptotic cells in VERU-111 treatment. (D) Effect of Protein levels of Bax, Bcl-2, Bad and Bcl-xl in Panc-1(i) and AsPC-1(ii) cells in response to VERU-111 treatment for 24 h as determined by Western blot analysis. (E) Effect of VERU-111 on protein levels of pro-caspase-3 and 9, cleaved caspase-3 and 9 and PARP cleavage in Panc-1(i) and AsPC-1(ii) cells as determined by Western blot analysis. (F) Effect of VERU-111 alone or in combination with caspase inhibitor Z-VAD-FMK on protein level of pro-caspase-3 and 9, cleaved caspase-3 and 9 and PARP cleavage in Panc-1(i) and AsPC-1(ii) cells as determined by Western blot analysis. (G) Effect of VERU-111 alone or in combination with caspase inhibitor Z-VAD-FMK on apoptosis induction analyzed by flow cytometry using Annexin V-7AAD Apoptosis kit. Data was acquired by using the Bio-RAD ZE5/Evererst Software v2.1 and analyzed using FlowJo v.10.3. Asterisk (*) denotes the significant value p < 0.05