Figure 5.

Effect of cytostatic concentrations of MF on DNA synthesis in OV2008 cells studied 10 days after being exposed to a lethal concentration of CDDP for 1 h. Cells were seeded and allowed to attach to the plate surface for 48 h, and exposed to vehicle (0.9 % NaCl) or 20 µM CDDP for 1 h. The treatment was then removed and fresh media without or with 5, 10, or 20 µM MF was replaced every 2 days. Ten days after CDDP exposure, the cells were pulsed with 10 µM BrdUrd for 6 h, washed with PBS, fixed with 4% paraformaldehyde, and subjected to immunocytochemistry using an anti-BrdUrd antibody (a). BrdUrd positive nuclei were quantified using a microscope with 400 X magnification and at least 500 cells were counted for each sample (b). The arrow indicates a scattered BrdUrd positive cell in the culture receiving CDDP plus 20 µM MF. Untreated cells in their logarithmic phase of growth, and cells that received only 20 µM MF for 10 days were also included in the study. The experiment was done in triplicates and the results are expressed as percentage of BrdUrd positive cells. Scale bar = 20 µm. Bars, mean ± SEM. ^*, P < 0.05 compared to untreated; ^†, P < 0.05 compared to CDDP.