Research Paper Volume 12, Issue 18 pp 17990—18007

Figure 5. Regulatory effects of minocycline on p53 deacetylation and PIAS1 degradation. (A) BV-2 microglial cells were stimulated with minocycline (20 μM) for the indicated time periods. The expression levels of lysine 379 acetylated p53 and PIAS1 were determined by western blot analysis. (B, C) Cells were transfected with siRNA against SIRT1 or control for 24 h, and then treated with minocycline for another 24 h. (D) After treatment with EX527 (10 μM) for 30 min, the cells were treated with minocycline for another 24 h. Whole-cell lysate proteins were extracted and subjected to western blot analysis to assess acetyl-p53, PIAS1, and SIRT1 expression. Similar results were obtained from at least three independent experiments.