Cell-cycle effects of 5-FU and LY294002. SNU-719 cells were incubated with 5-FU (10 μM) or LY294002 (20 μM), in single or combined treatments, for 72 h. (A) Cell-cycle distributions were measured by flow cytometry. The results are expressed as the mean percentages of cells in the different phases of the cell cycle in each sample (sum to 100%) in three independent experiments. (B) Western blot analysis of retinoblastoma 1 (RB1), cyclin D3, CDK2, cyclin A, and CDK4 was performed using the corresponding antibodies to confirm the phase-specific cell-cycle effects of 5-FU and LY294002 alone or in combination, as shown in (A). Representative immunoblots are shown from three independent experiments. Actin on the same immunoblot was used as the loading control.