Figure 2. Accumulation of HMGCR protein in tissues of WT and Ubiad1^Ki/Ki mice with C57BL/6 genetic background.

(A and B) Eight to nine-week old male WT, Ubiad1^WT/Ki, and Ubiad1^Ki/Ki littermates (six mice/group) were fed an ad libitum chow diet prior to study. Aliquots of membrane (Memb.) and nuclear extract (N.E.) fractions from homogenized livers, enucleated eyes, kidneys, brains, testes, and spleens (23–50 µg of total protein/lane) were analyzed by immunoblot using antibodies against the indicated proteins. The asterisk indicates a non-specific cross-reactive band observed in the anti-HMGCR immunoblot from brain and pancreas. Although shown in separate panels, LSD-1 serves as a loading control for the nuclear SREBP-1 and SREBP-2 immunoblots. In (B), the amount of HMGCR protein in the indicated tissues from Ubiad1^Ki/Ki mice was determined by quantifying the band corresponding to HMGCR using Image J software. (C) For mRNA analysis, equal amounts of RNA from the indicated tissue of individual mice were subjected to quantitative real-time RT-PCR using primers against the Hmgcr mRNA and cyclophilin mRNA as an invariant control. Error bars, S.E.

Figure 2—figure supplement 1. Accumulation of HMGCR protein in eyes and livers of WT and Ubiad1^Ki/Ki mice.

Female WT, Ubiad1^WT/Ki, and Ubiad1^Ki/Ki littermates of animals used in Figure 2 (six mice/group, 8–9 weeks of age) were fed an ad libitum chow diet prior to study. Aliquots of membrane (Memb.) and nuclear extract (N.E.) fractions from homogenized livers (A) and enucleated eyes (B) (50–80 µg of total protein/lane) were analyzed by immunoblot using antibodies against the indicated proteins. Although shown in separate panels, LSD-1 serves as a loading control for the nuclear SREBP-1 and SREBP-2 immunoblots in (A).