Fig. 3
H89 inhibited LMP1-promoted CNE1 cell proliferation. (a) CNE1 cells were transfected with pcDNA3.0 or pcDNA3.0-LMP1. H89 was added to the culture medium at the concentration indicated every 12 h after transfection. The expression of phosphorylated MSK1 at Thr581 was detected by Western blotting. β-actin was used as loading control. (b) CNE1G and CNE1GL cells were seeded in 96-well plates and incubated with different concentrations of H89 for 24 h or 48 h. Cell proliferation was estimated using CCK-8 assay. Data are presented as mean ± SD for three independent experiments. *P < 0.05; **P < 0.001, compared with CNE1GL cells untreated with H89. (c) CNE1GL cells were incubated with different concentrations of H89 for 2 weeks, and then the ability of colony formation was evaluated. *P < 0.01; **P < 0.005, compared with CNE1GL cells untreated with H89. (d) The effect of H89 on LMP1-induced cell cycle distributions was analyzed by flow cytometric assay. Data are expressed as the percentage of cells in G0/G1, S, or G2/M phase. *P < 0.05; **P < 0.005, compared with LMP1-transfected CNE1 cells untreated with H89