R-etodolac induces down-regulation of cyclin D1 expression. (A) U266 cells were cultured with control media (□) or 1.0 mM R-etodolac (▪) at the indicated times. Assessed by trypan blue exclusion, the percentage of cell growth is shown in the bar graph; and the percentage of cell viability is depicted by line. Data represent mean (± SD) of quadruplicate cultures. (B) U266 cells were cultured with or without R-etodolac (1.0 mM) for the indicated times. Total cell lysates were subjected to Western blotting using anti-cyclin D1 and -α-tubulin Abs. (C) Cell-cycle profile was assessed by propidium iodide (PI) staining and flow cytometric analysis. The percentage on each panel indicates the percentage of cells in the sub-G1 region (horizontal bar).