Figure 5. Regulation of HMGCR in livers of cholesterol-fed WT, Ubiad1^Ki/Ki, and Hmgcr^Ki/Ki mice.

Male mice (12–13 weeks of age, five mice/group) were fed an ad libitum chow diet supplemented with the indicated amount of cholesterol for 5 days. Aliquots of membrane (Memb.) and nuclear extract (N.E.) fractions from homogenized livers (A and C) or enucleated eyes (B) (70 µg protein/lane) were analyzed by immunoblot analysis with antibodies against the indicated proteins as described in the legend to Figure 1. The asterisk denotes a nonspecific band observed in the nuclear SREBP-2 immunoblot. (D) For mRNA analysis, equal amounts of RNA from livers of mice were subjected to quantitative real-time RT-PCR using primers against the indicated mRNAs and cyclophilin mRNA as an invariant control. Error bars, S.E. Pcsk9, proprotein convertase subtilisin/kexin type 9.

Figure 5—figure supplement 1. Effect of dietary cholesterol on expression of mRNAs encoding components of the Scap-SREBP pathway in livers of WT and Ubiad1 knock-in mice.

Total RNA from livers of mice used in Figure 5A (5 mice/group) was separately isolated. Equal amounts of RNA from the individual mice were subjected to quantitative real-time RT-PCR using primers against the indicated gene; cyclophilin mRNA was used as an invariant control. Each value represents the amount of mRNA relative to that in WT mice fed a chow diet, which was arbitrarily defined as 1. Bars, mean ± S.E. (error bars) of data from five mice. ApoE, apolipoprotein E; Acat-1, acyl-coenyzme A:cholesterol acyltransferase-1.