(A) Hela cells were treated with Ary at the indicated concentration for 24 h, and phos-ERK1/2 and ERK1/2 were detected in the treated cells using Western-blotting. (B) The untreated cells (a, b, c, d) and the treated cells (e, f, g, h) were stained with ERK1/2 mouse mAb (green), phosphorylation of ERK1/2 rabbit mAb (red) and DAPI, respectively. ERK1/2 and phos-ERK1/2 in treated cells were observed under Confocal microscopy. (a, e), DAPI; (b, f), ERK1/2; (c, g), phos- ERK1/2. (C) Hela cells were pretreated with U0126 at 10 μM for 2 h, then treated with 2.5 μg/ mL Ary treatment for 24 h. The expression ERK1/2, phos-ERK1/2 and CyclinA2 proteins were detected using Western-blotting. (D) The treated cells were analyzed by flow cytometry. (a), blank; (b), 2.5 μg/mL; (c), U0126; (d), U0126 and Ary. (E) Colony formation ability of the treated cells was determined using soft agar colony formation assay, and the inhibition rates of colony formation were calculated. Data were shown as the mean ± SD of three independent experiments by analysis of Student's t test. *P < 0.05, **P < 0.01.