Hsp90 Blockers Inhibit Adipocyte Differentiation and Fat Mass Accumulation
Figure 2
Hsp90 inhibition decreases the adipogenic transcriptional program without any effect on cytotoxicity.
(A) Primary preadipocytes derived from white adipose tissue (WAT) of mouse gonadal fat were cultured for 10 days with 17-AAG (300 nM) or DMSO (Control) and visualized by microscopy 40X (scale bar: 50 μm). (B) Mature 3T3-L1 were cultured for 2 days with or without 17-AAG (100 nM) and tested for an additional 2 h for lipolysis in response to 1 μM of isoproterenol. (C) 3T3-L1 cells were induced to differentiation in presence of 17-AAG for 2 days then tested for [3H]-Deoxy-D-glucose uptake for 10 min. (D) 3T3-L1 preadipocytes were induced to differentiation in presence or absence of 17-AAG for 10 days. The abundance of CEBPα, PPARγ, LPL and Glut4 mRNA in 3T3-L1 cells was measured by quantitative RT-PCR. Given are means relative to GAPDH of 3 experiments performed in duplicates ± SEM, **p<0.01. (E) Cell lysates were analyzed by immunoblotting using antibodies against Hsp90, PPARγ, CEBPα and actin as a loading control. (F) The protein expression levels were quantified and values are given as mean ± SEM (n = 6), **p<0.01.