Figure 4.

Effect of 5-FU + BA combination on apoptosis in RMS cell lines. A. The RMS cells were treated for 48 h with BA and combination of 5-FU + BA in the presence or absence of 50 μM of the zVAD.fmk. Apoptosis-specific DNA fragmentation was determined by FACS analysis of propidium iodide-stained nuclei; 100 μg/ml cycloheximide (CHX) was used as a positive control for apoptosis induction. B. The RMS-13 cells were treated for 48 h with BA and combination of 5-FU + BA in the absence or presence of zVAD.fmk. Expression of caspase 3 and β-actin was detected by western blot analysis. 100 μg/ml cycloheximide (CHX) was used as a positive control for apoptosis induction. C. The RMS-13 cells were treated for 24 and 48 h with BA and combination of 5-FU + BA. Mitochondria were isolated, lysed, and the expression of cytochrome c and COX4 in the mitochondrial and cytosolic fraction was detected by western blot analysis