FIG. 3.

SirT1 requires PGC-1α to induce the expression of oxidative stress protection genes. (A, B) BAEC were coinfected o/n with Ad-SirT1 and Ad-shPGC-1α, or the respective control adenoviruses. 12 h postinfection, cells were deprived of serum o/n and harvested. Expression of the oxidative stress protection genes was analyzed by qRT-PCR (A) and Western Blot (B). (C, D). BAEC whole cell extracts were subjected to immunoprecipitation (IP) with a specific PGC-1α antibody or normal IgG, as a control, and analyzed by western blot as indicated. Input indicates a 1/10 of whole cell extract not subjected to immunoprecipitation. (C) BAEC were infected o/n with Ad-PGC-1α and 8 h postinfection were treated with 10 mM nicotinamide for 15 h. (D) BAEC were coinfected o/n with Ad-SirT1 and Ad-PGC-1α, or the respective control adenoviruses. 24 h postinfection cells were deprived of serum o/n, and then treated with 50 μM H₂O₂ or vehicle for 4 h. (E) Confluent BAEC were grown in galactose media and in the absence of serum for 48 h. Whole cell extracts were subjected to immunoprecipitation (IP) with specific PGC-1α or SirT1 antibody or normal IgG, as a control, and analyzed by western blot as indicated. Data are from ≥3 independent experiments. Data are means+SD (*) p≤0.05 versus control. The blots presented correspond to a representative experiment.