Fig. 1
From: RORα controls hepatic lipid homeostasis via negative regulation of PPARγ transcriptional network
Liver-specific Rorα deleted mice are susceptible to diet-induced obesity. a Schematic representation of the Rorα gene-targeting strategy, including a map of the RORα exon 4 and 5 allele (yellow box) and the targeting vector with loxP sites (red arrowhead), FRT sites (blue box), and puromycin selection gene (green box). Bg: BglII, RI: EcoRI, Bh: BamHI, Kp: KpnI, Sp: SpeI. b Southern blot analysis to screen correctly targeted Rorα + /puro ES cell clones. For BamHI digestion, the bands representing WT and mutant alleles were 9.0 kb and 6.8 kb, respectively. PCR analyses with genomic DNA extracted from tail of WT, RORαf/+, Alb; RORαf/+, RORαf/f and Alb; RORαf/f mice are shown. PCR were performed to amplify the cre (top), floxed and deleted allele (bottom). c, d mRNA expression level of RORα in liver extract c and primary hepatocyte d from RORαf/f and RORαLKO mice. Expression was normalized to 18 s rRNA expression. e, f Protein expression level of RORα in liver extract e and primary hepatocyte f. g Body weight change in RORαf/f and RORαLKO mice fed CD or HFD for 10 weeks (n = 9–12/group). Statistical analysis was performed using Student’s unpaired t-test. *P < 0.05, RORαf/f vs. RORαLKO, HFD. h, i RORαf/f and RORαLKO mice were fed with HFD for 10 weeks. h Body composition analysis of RORαf/f and RORαLKO mice (n = 6/group). i Macroscopic views of RORαf/f and RORαLKO mice. j Adipose tissues weight of RORαf/f and RORαLKO mice (n = 6–7/group). k Representative image of epidydimal white adipose tissue (eWAT) from RORαf/f and RORαLKO mice stained with hematoxylin and eosin. Scale bar, 100 μm. l, m Expression levels of inflammatory cytokine genes in eWAT extract l or thermogenesis genes in BAT extract m from RORαf/f and RORαLKO mice (n = 4–5 per group) as determined by qRT-PCR. Expression was normalized to L32 expression. n Metabolic cage studies were performed in RORαf/f and RORαLKO mice (n = 5–6 mice/group). O2 consumption (VO2), CO2 production (VCO2) and energy expenditure were represented (left to right). Statistical analysis was performed using Student’s unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001, NS, Non-Significant. Data expressed as mean ± s.e.m
