FIGURE 3.

ANXA2 knockdown causes G₂ arrest. A small interfering RNA (siANXA2) was used to silence ANXA2 in A549, H460, and SiHa cells. A non-targeting siRNA (scramble) was used as a negative control. A and B, the cell cycle stages (G₁, S, and G₂/M) of siANXA2-transfected cells were determined using PI staining followed by flow cytometry 4 days post-transfection. One representative data set and the percentage of cells in the G₂/M phase are shown as the means ± S.D. from three individual experiments with triplicate cultures. ***, p < 0.001 compared with scramble. C, the M phase cells were determined by analysis of DNA content (PI) and MPM2 staining 4 days post-transfection with or without nocodazole treatment given at 3 days post-transfection. Nocodazole treatment in A549 cells for 24 h was used as a positive control. The percentages of G₂ and M phase (MPM2-positive) cells are shown as the means ± S.D. from three individual experiments with triplicate cultures. ***, p < 0.001 compared with scramble. D, Western blotting was used to determine the expression of ANXA2, cyclin B1, and Cdc2 for the indicated times. β-Actin was the internal control. The data shown are representative of three individual experiments.