Fig. 3.

SIRT6 negatively regulates AKT signaling. a HEK293T cells were transfected with Flag or Flag-SIRT6 for 48 h, and then immunoprecipitated with anti-SIRT6. IgG was used as a negative control. b HEK293T cells were transfected with shCtrl or shSIRT6 for 48 h, cells lysates was immunopreciptated with anti-AKT, acetylated AKT was performed using anti-acetylated-lysine. c ChIP analysis using anti-SIRT6 to detect SIRT6 at the promoter of AKT in SIRT6 overexpression HEK293T cells. The data are expressed as mean ± SEM (n = 3). ***P < 0.001 compared with His. d ChIP analysis using anti-H3K9ac or anti-H3K56ac to detect histone at the promoter of AKT in SIRT6 overexpression HT-22 cells. The data are expressed as mean ± SEM (n = 4). ***P < 0.001 compared with Myc-H3 + SIRT6. e Immunoprecipitation analysis using anti-Myc to detect acetylated-histone in SIRT6 overexpression HEK293T cells. f, g SIRT6 condensed chromatin and inhibited AKT signaling by deacetylating H3K9 and H3K56. Conversely, suppression of SIRT6 relaxed chromatin and promoted AKT signaling by increasing H3K9 and H3K56 acetylation