Fig. 5.

HSC70 participates in AICAR-induced FSP27 degradation. A and B: 3T3-L1 adipocytes were transfected with control or HSC70-specific siRNA. After 3 days, cells were treated with 10 μg/ml cycloheximide (CHX) for the indicated times. Endogenous FSP27 was detected by IB with anti-FSP27 antibody. C–F: 3T3-L1 adipocytes were transfected with control or HSC70-specific siRNA for 1 day and then infected with Ad-FSP27-FLAG. Two more days later, cells were treated with 10 μg/ml CHX (C and D) or 2 mM AICAR (E and F). FSP27-FLAG was detected by anti-FLAG immunoblotting. The levels of endogenous or ectopic FSP27 were quantified by the densitometric measurement, using β-actin as a loading control (B, D, and F). Data are presented as the mean ± SD of 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. Ctrl knockdown.