Figure 2

Analysis of cell cycle effects and cell apoptosis of Rm-HE in Jurkat cells. (A) Cell cycle analysis of Jurkat cells treated with Retama monosperma Hexanic Extract (Rm-HE) by flow cytometry. Jurkat cells (4 × 10 cells/ml) were incubated with 40 μg/ml of Rm-HE as indicated for 24 h and 48 h. Cells were harvested and their DNA content analysed by flow cytometry as described in Materials and Methods. The cell cycle distribution is shown for each experimental condition. (B) The graph summarizes the percentage of each phase in control, 24 h and 48 h-treated cells, respectively. (C) Effect of Rm-HE on pH2A.X levels. Cells (4 × 10cells /ml) were treated with 40 μg/ml Rm-HE for 0, 2, 4, 8 and 16 h. p-H2A.X levels in cellular extracts were detected by immunoblot with specific antibodies. Tubulin was used as an internal control. (D) Effect of Rm-HE in Jurkat cell apoptosis analyzed by flow cytometry. Cells were treated with 40 μg/ml of Rm-HE for 12 h and 24 h. The x-axis shows Annexin V-FITC staining and y-axis indicates Propidium iodide staining; Lower left (LL) quadrant: viable cells; lower right (LR) quadrant: early apoptotic cells; upper left (UL) quadrant: necrotic cells, upper right (UR) quadrant: late apoptotic cells.