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. Author manuscript; available in PMC: 2011 Nov 23.

Published in final edited form as: Circulation. 2010 Nov 8;122(21):2170–2182. doi: 10.1161/CIRCULATIONAHA.110.958033

Cultured myocytes were transduced with adenovirus harboring LacZ (control, Ad-LacZ), wild type Sirt1 (Ad-Sirt1-WT), shRNA-Sirt1 (Ad-sh-Sirt1) or tet-inducible p300. Expression of p300 was induced by co-transduction with adenovirus harboring tTA without doxycycline. Forty-eight hours after transduction, cells were harvested. (A) Expression of FoxO1, acetylated FoxO1 and tubulin (internal control) was evaluated with immunoblot analyses. (B,C) The relative expression of FoxO1 and acetylated FoxO1 was evaluated by densitometric analyses. The expression level of FoxO1 and acetylated FoxO1 in Ad-LacZ transduced myocytes is expressed as 1. (D,E) Myocytes were transduced with adenovirus harboring LacZ, wild type FoxO1 (Ad-FoxO1-WT) and shRNA-FoxO1 (Ad-sh-FoxO1). (D) Expression of MnSOD and tubulin (internal control) was evaluated with immunoblot analyses. (E) The relative expression of MnSOD was evaluated by densitometric analyses. The expression level of MnSOD in Ad-LacZ transduced myocytes is expressed as 1. (F) Myocytes were transfected with 3xIRS-Luc and then transduced with Ad-LacZ or Ad-Sirt1 (5 MOI). Ad-Sirt1 transfected myocytes were treated with or without sirtinol (10 μM). Experiments were conducted three times.

Cultured myocytes were transduced with adenovirus harboring LacZ (control, Ad-LacZ), wild type Sirt1 (Ad-Sirt1-WT), shRNA-Sirt1 (Ad-sh-Sirt1) or tet-inducible p300. Expression of p300 was induced by co-transduction with adenovirus harboring tTA without doxycycline. Forty-eight hours after transduction, cells were harvested. (A) Expression of FoxO1, acetylated FoxO1 and tubulin (internal control) was evaluated with immunoblot analyses. (B,C) The relative expression of FoxO1 and acetylated FoxO1 was evaluated by densitometric analyses. The expression level of FoxO1 and acetylated FoxO1 in Ad-LacZ transduced myocytes is expressed as 1. (D,E) Myocytes were transduced with adenovirus harboring LacZ, wild type FoxO1 (Ad-FoxO1-WT) and shRNA-FoxO1 (Ad-sh-FoxO1). (D) Expression of MnSOD and tubulin (internal control) was evaluated with immunoblot analyses. (E) The relative expression of MnSOD was evaluated by densitometric analyses. The expression level of MnSOD in Ad-LacZ transduced myocytes is expressed as 1. (F) Myocytes were transfected with 3xIRS-Luc and then transduced with Ad-LacZ or Ad-Sirt1 (5 MOI). Ad-Sirt1 transfected myocytes were treated with or without sirtinol (10 μM). Experiments were conducted three times.