Figure 3.

HSPB1 is induced by Ac2-26 in microglia. RT-PCR analysis of total RNA isolated from rat brain tissues, neurons, astrocytes and microglial cells revealed the presence of sHSP transcripts (A,B). BV-2 cells were treated with Ac2-26 (10 μM) before they were continuously exposed to oxygen glucose deprivation (OGD) for 1 h and reperfusion for 24 h. The mRNA expression of HSPB1 was assessed by RT-PCR (C), and the protein levels were detected by Western blotting (D) as indicated. (E,F) Microglia were treated with NCM or Ac2-26 (10 μM) for 24 h. Protein levels were assessed by Western blotting as indicated. The data are representative of at least three independent experiments and shown as the mean ± SEM. All data were analyzed using one-way ANOVA with Tukey’s multiple comparison tests; Ac2-26 induced HSPB1 protein expression (E) was analyzed using unpaired Student’s t test. *P < 0.05; **P < 0.01; ***P < 0.001 vs. control. (G) Protein cell lysates from HeLa cells transfected with GFP-ANXA1 were immunoprecipitated with anti-GFP mAb or mouse IgG and immunoblotted using an anti-HSPB1 mAb. This experiment was repeated at least three times.