Fig. 2

ONC201 induced dose-dependent growth inhibition in EC cell lines. Four endometrial EC cell lines, Ishikawa, ECC-1, HEC1A and KLE, were treated with various doses of ONC201 for 72 h. Cell proliferation was determined by MTT assay. Relative survival was determined by dividing the number of remaining ONC201 treated cells by the number of remaining viable DMSO (control). Representative dose-response curves and IC50 values are shown (a). The effect of ONC201 on the colony forming ability in ECC-1 and KLE cells was assessed through a colony-formation assay (b). The ECC-1 and KLE cells displayed distinct morphological alterations after 48 h treatment with ONC201 (c). Western blot analysis was performed to detect the expression of DRD2 in the four EC cell lines (d) and alternations of DRD2 and DRD5 after ONC201 treatment for 24 h in ECC-1 and KLE cell lines (e). The ECC-1 and KLE cells were cultured with various concentrations of ONC201 for 24 h followed by Western blot analyses to determine the expression of phosphorylation of AKT and S6. The results demonstrate that ONC201 inhibited the mTOR pathway in both cell lines (f). The levels of β-actin or α-tubulin served as protein loading controls. Data are representative of one of three independent experiments