Ovatodiolide inhibits RCC cell viability and induces apoptosis. (a) The effect of ovatodiolide on cell viability was evaluated in 786-O, Caki-1, and HK-2 cells by MTT assay at various times. The IC50 at 48 hr was calculated, and the relative viability was the percentage of MTT absorbance of ovatodiolide to DMSO-treated cells. Data are mean ± SD of triplicate experiments. (b) Flow cytometry of the effect of ovatodiolide on cell cycle distribution. 786-O and Caki-1 cell lines were treated with 40 μM ovatodiolide for 48 hr, and sub-G1 and G2/M populations were measured. Data are mean ± SD of triplicate experiments. (c) Western blot analysis of protein levels of cleaved caspase 8 and 9, and apoptotic proteins Bax, Bid and, PUMA, and antiapoptotic proteins Bcl-2, Bcl-xL, and survivin in 786-O and Caki-1 cell lines treated with ovatodiolide for 24 or 48 hr.