Fig. 3

ONC201 induced cell cycle G1 arrest and apoptosis in EC cells. The ECC-1 and KLE cells were treated with various doses of ONC201 for 36 h. Cellometer analysis showed ONC201 induced cell cycle arrest in G1 phase in both cell lines (a). Cell lysates from the ECC-1 and KLE cell lines were subjected to Western blotting analysis for CDK4 and Cyclin D1 after treatment ONC201 for 24 h (b). Apoptosis was determined by Annexin V/ PI analyses after 24 h of treatment with ONC201 in ECC-1 and KLE cells. The profiles of Annexin V-FITC/PI staining are presented. The percentage of early apoptotic cells is located in the lower right quadrant (c). ECC-1 and KLE cells were cultured with different concentrations of ONC201 for 24 h, and then subjected to Western blot analysis on the levels of anti-apoptotic proteins. ONC201 reduced BCL-2 and MCL-1 expression in both cell lines (d). Data are representative of one of three repeats