Fig. 5

Sirt6 directly interacted with and deacetylates PGC-1α. a, b HiBEC were transfected with pcDNA3-HA or Sirt6 for 48 h, before harvest, then treated with 1 mM AICAR or 5 μM Comp C for 12 h, cells were harvested for western blot and RT-qPCR assay. c HiBEC were transfected with ‒ 1000/ + 1-LUC PGC-1α construct (200 ng) plus pRL-TK-Renilla (10 ng), pcDNA3-HA (200 ng), Sirt6 (200 ng), AMPKα1 (200 ng) and AMPKα1 (DN) (200 ng), treated with or without 1 mM AICAR or 5 μM Comp C for 12 h. PGC-1α promoter driven luciferase activity were normalized to Renilla luciferase activity, presented as mean ± SEM. d HiBEC were transfected with vector control, Sirt6 or PGC-1α for 48 h, total lysates were harvest, immunoprecipitated with antibody to Sirt6 or PGC-1α, and immunoblotted with antibodies to PGC-1α and Sirt6. e Flag-Sirt6 was co-expressed with HA-tagged PGC-1α, immunoprecipitated with antibody to Flag or HA, and immunoblotted with antibodies to HA or Flag. Cell extracts were also immunoblotted with antibodies to HA, Flag or GAPDH. f HiBEC cells were transfected with Sirt6 or si-Sirt6 for 48 h, total cell lysates were immunoprecipitated with antibody to PGC-1α then immunoblotted with antibody to Ac-K. *P < 0.05