Figure 6.

SIRT1 is dose- and time-dependently shuttled out of the nucleus by oxidants. A) BEAS-2B cells were treated with CSE (0.5–1.5%) or H₂O₂ (150 μM) for 1 h. B) BEAS-2B cells were treated with CSE (0.5%) for 30 min, 1 h, and 6 h. Cells were fixed with 4% paraformaldehyde and used for immunostaining. SIRT1 was visualized by using goat anti-rabbit secondary antibody conjugated to Alexa Fluor 594 dye. Nuclei were stained with Hoechst 33342 dye. Untreated cells that received Hoechst 33342 and secondary antibody but no SIRT1 primary antibody was used as a negative control. Arrows denote SIRT1. Images represent 3 separate experiments (n=3).