FIGURE 7.

OA-NO₂ induces HSP70 expression via HSF1. A and B, HUVECs were transfected with 100 nm control or HSF1 siRNA. 72 h after transfection, cells were treated with 3 μm OA-NO₂ for 4 h (A) or 8 h (B) or with heat shock for 30 min. The expression levels of HSF1 and HSP70 (HSPA1A) were analyzed with quantitative real time PCR (A) or Western blot (B). Values are expressed as mean ± S.E., n = 3. *, p < 0.05 versus control. Western blots are representative of three independent experiments. C, HUVECs were treated with 5 μm OA-NO₂ for 4 h. The binding of HSF1 to the promoter region of HSP70 was determined with chromatin immunoprecipitation using β-actin as a control. D, HUVECs were transfected with 50 nm control or Nrf2 siRNA. 72 h after transfection, cells were treated with OA-NO₂ for 4 h. The binding of HSF1 to the promoter region of HSP70 was determined with chromatin immunoprecipitation using β-actin as a control.