Fig. 6.

Effect of MIG12 on de novo FA synthesis and TG accumulation in rat primary hepatocytes. A and B, Hepatocytes were infected with 20 MOI of adenoviruses for the expression of β-gal (Ad-Con) or mMIG12 (Ad-MIG12) as described in Materials and Methods. At 24 (A and B) and 48 h (B) after infection, cells were placed in fresh medium B containing the vehicle (DMSO) or 10 μm T0901317. Forty-eight hours after infection, cells were treated with [¹⁴C]-acetate, and incorporation of ¹⁴C into newly synthesized FAs was determined as described in Materials and Methods (A). Seventy-two hours after infection, intracellular TG levels were determined as described in Materials and Methods (B). C–E, Hepatocytes were transfected with 150 pmol of control siRNA (siCon) or siRNA targeting MIG12 (siMIG12) as described in Materials and Methods. Twenty-four hours after transfection, cells were placed in fresh medium B containing vehicle (DMSO) or 10 μm T0901317. Seventy-two hours after transfection, total RNA was extracted, and real-time PCR analysis was performed. Relative mRNA levels were obtained after normalizing to cyclophilin mRNA levels. mRNA levels of MIG12 transfected with siCon in the absence of T0901317 are represented as 1 (C). Forty-eight hours after transfection, cells were treated with [¹⁴C]-acetate, and ¹⁴C incorporation into newly synthesized FAs was determined as described in Materials and Methods (D). Seventy-two hours after transfection, intracellular TG levels were determined as described in Materials and Methods (E). All results are expressed as mean ± sd; experiments were performed in triplicate. Similar results were obtained in three independent experiments. *, P < 0.05; **, P < 0.01.