Figure 2

Effects of inhibiting PI3 K/PI3K/Akt or Raf/MEK/Erk on growth of RoSH2 cells. RoSH2 cells were pre-labeled with CFDA, cultured on gelatin-coated plates for 24 hours. One quarter of the plates were not treated with any inhibitor (N), one quarter with 10 μM Ras inhibitor, FTase inhibitor III (FTI), one quarter with 50 μM MEK inhibitor PD98059 (PD), and the last quarter with LY 294002 (LY). Each of the media with the appropriate inhibitors was replaced every twenty-four hours. At 0, 24, 48 and 72 hours after plating, cells were harvested and were assayed for cellular fluorescence by flow cytometry. The number of cell cycles for every 24 hours was calculated as described in Materials and Methods. b) Effects of FTI and PD98059 on cell numbers. Twenty-four hours after treatment with FTI, PD98059 or a combination of both drugs, the cell numbers were counted and normalized against that of untreated cells. Cell number at time 0 was designated as one.