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. 2014 Mar 20;289(19):13132–13141. doi: 10.1074/jbc.M113.520734

FIGURE 2.

FIGURE 2.

Inhibition of class III HDACs, the sirtuins, by nicotinamide induces acetylation of S6K1. A, S6K1 but not S6K2 acetylation (Ac-S6K) is induced by NT treatment. COS-7 cells were transfected with HA-S6K1 or HA-S6K2 plasmid. 48 h after transfection, the cells were treated with NT for 3 h. HA-S6Ks were IPed, and levels of acetylation were analyzed by immunoblotting with anti-acetyl-lysine antibody. B and C, NAM, but not TSA, induces S6K1 acetylation. HA-S6K1 was expressed in COS-7 (B) or HEK293T cells (C). Cells were treated with NAM, TSA, or NT for 3 h. Levels of S6K1 acetylation were monitored as in A. D and E, acetylation of endogenous S6K1 was enhanced by NT treatment. HEK293T (D) or COS-7 cells (E) were treated with or without NT for 3 h. Endogenous S6K1 was IPed, and levels of S6K1 acetylation were monitored as in A. Abs, antibodies; ctr IgG, control IgG.