FIGURE 4.

CDDO-Me down-regulates ACC activity in primary murine hepatocytes. Treatment of hepatocytes with CDDO-Me suppresses lipogenic genes in primary murine hepatocytes. A and B, Cyber green-based real time RT-PCR was used to measure relative mRNAs level of FAS, SREBP-1c, and HMGR (A) and ACC1 (B). In each case, the data were normalized to the expression (Exp.) level of actin and are expressed as 2^−ΔCT, and then the relative mRNA expression level was calculated. Deactivation of ACC (dephosphorylation) and ACC protein level were checked by Western blot using anti-phospho-ACC and anti-ACC antibody in hepatocytes (C) and liver samples (D). E, CDDO-Me (solid bar, 100 nm) and AICAR (hatched bar, 250 μm) significantly inactivate ACC activity in mouse primary hepatocytes in comparison with controls (empty bars) when treated for 7 h. F, palmitate oxidation in hepatocytes plated in 60-mm tissue culture plates. ¹⁴CO₂ were trapped using hyamine hydroxide soaked blotting paper in each cover. The amount of CO₂ produced was then normalized using total protein and expressed as a percentage of production of ¹⁴CO₂ (fold over basal). The corresponding bar graph shows the quantification of the blot. All of the values are shown as the means ± S.D. *, p < 0.05, versus control.