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

Figure 5. SHP-1 is involved in andrographolide-induced PP2A dephosphorylation in rat VSMCs.

Figure 5

(a)Cells were treated with 50 μM andrographolide for the indicated time periods. Cells were then harvested and PP2A-C demethylation and Tyr-307 phosphorylation were examined by immunoblotting. Compiled results are shown at the bottom of the chart. Each column represents the mean ± S.E.M. of four independent experiments. The full-length blots are presented in Supplementary Figure 4a. Cells were pretreated with vehicle or 10 μM SSG for 30 min followed by the treatment with 50 μM andrographolide for another 30 min (b) or 48 h (c). The extent of PP2A-C Tyr-307 phosphorylation was them examined by immunoblotting (b). 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 4b. Cell viability was determined by MTT assay (c). Each column represents the mean ± S.E.M. of four independent experiments. (d) Cells were transiently transfected with shp-1 siRNA for 48 h followed by the treatment with andrographolide for another 30 min. The extent of PP2A-C Tyr-307 phosphorylation was them examined by immunoblotting. Compiled results are shown at the bottom of the chart. Each column represents the mean ± S.E.M. of four independent experiments. The full-length blot is presented in Supplementary Figure 4c. (e) Cells were transiently transfected with shp-1 siRNA for 48 h followed by the treatment with andrographolide for another 48h. Cell viability was then determined by MTT assay. Each column represents the mean ± S.E.M. of four independent experiments. (f) Cells were treated as described in (d). The extent of SHP-1 was then examined by immunoblotting. Compiled results are shown at the bottom of the chart. Each column represents the mean ± S.E.M. of four independent experiments. The full-length blot is presented in Supplementary Figure 4d. (*, P < 0.05, **, P < 0.01, and ***, P < 0.001, compared with the control group; and #, P < 0.05, and ###, P < 0.001, compared with the group treated with andrographolide alone).