Figure 3

Effects of Rm-HE on apoptosis induction and activation of caspases in Jurkat cells. (A) Effect of Retama monosperma Hexanic Extract on caspase activity induction on Jurkat cells. Jurkat cells were treated with Rm-HE (40 μg/ml) or 1 μM Doxorubicin (positive control) for 24 h and 48 h and caspase activity was measured as indicated in Materials and Methods. Results indicate the average fold increase ± S.E.M in caspase activity relative to untreated cells from three independent determinations performed in duplicate. (B) Effect of Retama monosperma Hexanic Extract on viability in the presence of a caspase inhibitor Jurkat cells were pre-incubated for 1 h with 5 μM Q-VD-OPh and Rm-HE (40 μg/ml) was added for 24 h. Doxorubicin was used as positive control. Cell viability is represented as a percentage relative to untreated cells, and data is means ± S.E.M. from three independent determinations performed in duplicate. (C) Effect of Retama monosperma Hexanic Extract on the expression and cleavage of caspases 8, 7, 3 and 9 in Jurkat cells. 4 × 10cells were treated with Rm-HE (40 μg/ml) for 0, 2, 4, 8 and 16 h. Total and cleaved caspase levels in cellular extracts were detected by immunoblot with specific antibodies. Tubulin was used as an internal control.