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. 2014 Jul 10;4:5651. doi: 10.1038/srep05651

Figure 3. Involvement of p38MAPK in andrographolide-induced p53 phosphorylation in rat VSMCs.

Figure 3

Cells were treated with 50 μM andrographolide for the indicated periods. The extent of p53 phosphorylation (a) and p38 phosphorylation (b) were then determined by immunoblotting. Compiled results are shown at the bottom of the chart. Each column represents the mean ± S.E.M. of at least three independent experiments. The full-length blots are presented in Supplementary Figure 2a and 2b (c) Cells were pretreated with vehicle, 10 μM SB203580, or 1 μM p38MAPK inhibitor III for 30 min followed by the treatment with 50 μM andrographolide for another 48 h. Cell viability was then determined by MTT assay. Each column represents the mean ± S.E.M. of four independent experiments. (d) Cells were transiently transfected with PG-13-luc and Renilla-luc for 48 h. After transfection, cells were pretreated with vehicle or SB203580 (10 μM) for 30 min followed by the treatment with andrographolide (50 μM) for another 24 h. The PG13-luciferase activity was then determined. Each column represents the mean ± S.E.M. of four independent experiments. (e) Cells were pretreated with vehicle or 10 μM SB203580 for 30 min followed by the treatment with 50 μM andrographolide for another 48 h. The extent of Bax level was then determined by immunoblotting. Compiled results are shown at the bottom of the chart. Each column represents the mean ± S.E.M. of three independent experiments. The full-length blot is presented in Supplementary Figure 2c. (*, P < 0.05, **, P < 0.01, and ***, P < 0.001, compared with the control group; and #, P < 0.05 and ##, P < 0.01, compared with the group treated with andrographolide alone).