Fig. 4.
The effects of 22-HC and ETYA on the cell-cycle status and gene regulation. (A) Cell-cycle analysis of RWPE-1 cells measured by FACS analysis. Cells were treated with ligand ED25 concentrations individually and in combination for 72h. Representative cell-cycle distributions are shown (upper panel), with subsequent quantification of three biological replicate populations (lower panel). Significant changes (determined by t-test) of cell population at any given cell cycle are noted, comparing additive results to those observed experimentally. (B) PPARγ candidate gene expression profile in RWPE-1 cells. Expression of candidate PPARγ-regulated genes (LXR-a, PTGS-2, TGFBRAP1, ALOX 5) at different time points of treatment (4, 12, 24h) determined via Q-RT–PCR. ETYA and 22-HC were dosed individually and in combination at ED25 concentrations and were compared with respective vehicle controls. Significant changes of gene expression between combination treatment and individual treatments (*ETYA, #22-HC) are noted.