Research Paper Volume 13, Issue 12 pp 16381—16403

Figure 8. Sirt1 mediates acacetin-induced Sirt6 activation by enhancing NAMPT and NAD⁺/NADH in H9C2 cardiac cells. (A) Western blots and relative levels of Sirt6 in cells transfected with control siRNA or Sirt6 siRNA in the absence or presence of 3 μM acacetin (Aca) for 72 h. (B) Western blots and relative levels of Sirt1 in cells as treated in A. (C) Western blots and relative levels of Sirt1 in cells transfected with control siRNA or Sirt1 siRNA in the absence or presence of 3 μM acacetin for 72 h. (D) Western blots and relative levels of Sirt6 in cells as treated in C (n = 4–6, ^**P < 0.01 vs. control siRNA, ^##P < 0.01 vs. control siRNA with acacetin). (E) Acacetin-induced increase of NAD⁺/NADH ratio was prevented by NAMPT inhibitor GNE-617 (50 nM). (F) Acacetin-induced increase of Sirt1 protein was abolished by 50 nM GNE-617. (G) Acacetin-induced increase of Sirt6 protein was abolished by 50 nM GNE-617. (n = 4–6, ^**P < 0.01 vs. control, ^##P < 0.01 vs. acacetin alone).