Figure 3.

SCF^Cdc4 targets phosphorylated PGC-1α for proteolysis. (A) HEK293T cells were cotransfected with the indicated alleles of PGC-1α and Cdc4γ. PGC-1α protein levels were determined by immunoblotting with anti-Flag antibody. (B,C) HEK293T cells were transfected with vector alone or Cdc4γ and different alleles of PGC-1α. Cyclohexamide (CHX) was added to cell cultures 48 h post-transfection, whole-cell extracts were prepared at the indicated times after CHX addition, and PGC-1α protein levels were analyzed by immunoblotting. GFP was cotransfected as a transfection and loading control.