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  • The effect of SIRT1 protein knock down on PGC-1α acetylation during skeletal muscle contraction.

The effect of SIRT1 protein knock down on PGC-1α acetylation during skeletal muscle contraction.

Journal of exercise nutrition & biochemistry (2015-01-08)
Dae Ryoung Park, Jeong Seok Kim, Chang Keun Kim
ABSTRACT

The purpose of this study was to investigate the effect of Sirtuin 1 (SIRT1) and General control nonderepressible 5 (GCN5) knock down on peroxisome proliferator- activated receptor gamma coactivator 1-alpha (PGC-1α) deacetylation during electrical stimulated skeletal muscle contraction. Skeletal muscle primary cell were isolated from C57BL/6 mice gastrocnemius and transfected lentiviral SIRT1 and GCN5 shRNA. Knock downed muscle cell were stimulated by electrical stimulation (1Hz, 3min) and collected for PGC-1α deceatylation assays. Immunoprecipitation performed for PGC-1α deacetylation, acetyl-lysine level was measured. Our resulted showed SIRT1 knock down not influenced to PGC-1α deacetylation during electrical stimulation induced muscle contraction while GCN5 knock down decreased PGC-1α deacetylation significantly (p<0.05). This study can be concluded that GCN5 is a critical factor for muscle contraction induced PGC-1α deacetylation.

MATERIALS
Product Number
Brand
Product Description

Millipore
Protein G–Agarose, lyophilized powder, Contains lactose stabilizers that must be removed prior to use.
Sigma-Aldrich
Anti-PGC-1 Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Anti-Sirt1(Sir2) Antibody, Upstate®, from rabbit