FIGURE 5.

Relative mRNA levels of selected genes involved in cholesterol and triglyceride metabolism in the livers of WT and Tm6sf2^−/− mice (A) and hepatic SREBP-1c cleavage (B). A, quantitative real-time PCR assays were performed to assess the relative levels of selected mRNAs in livers of the 13-week-old chow-fed male mice (5 mice/group) and in 11-week-old chow-fed female mice (6 mice/group) described in the legend to Fig. 3A. Expression levels were normalized to levels of 36B4 and expressed relative to levels of WT transcript. Values are means ± S.E. (error bars). The official gene symbols were used for all of the genes with the following exceptions: ATGL (adipose TG lipase), PNPLA2; LXRα, NR1H3; PGC-1α, PPARGC1A; L-PK, PKLR; PEPCK, PCK1; ChREBP, MLXIPL. B, SREBP-1c regulation in Tm6sf2 KO mice. Nuclear and membrane fractions were isolated from livers of 18-week-old refed male mice (n = 4) by ultracentrifugation as described under “Experimental Procedures.” Lysates from each mouse were pooled, and 40 μg of pooled protein was size-separated on SDS-10% polyacrylamide gels. Proteins were transferred to nitrocellulose membranes and blotted with rabbit anti-mouse mSREBP-1c antibody. The bands were visualized by ECL and quantified using a LI-COR Odyssey Fc imager. The membranes were then stripped and reblotted with antibodies against calnexin and LSD1. The experiment was repeated with 13-week-old females, and the results were similar. *, p < 0.05; ****, p < 0.0001.