Liver-specific disruption of PPARγ in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes
J. Clin. Invest. Kimihiko Matsusue, et al. 111:737 doi:10.1172/JCI17223 [Go to this article.]

Figure 1
Gene targeting and conditional deletion of exon 2 of the PPARγ gene. (a) Restriction maps of the wild-type allele, targeting vector, targeted allele, floxed allele, and null allele. The indicated 3′ probe was used to assess recombination events by Southern blot analysis. Open boxes represent exons and are numbered as indicated. PGK neomycin (PGK Neo) and thymidine kinase (TK) are positive and negative selection cassettes, respectively. Restriction sites: B, BamHI; E, EcoRI; S, SacI. (b) Southern blot analysis of BamHI-digested genomic DNA isolated from brain (B), liver (L), colon (C), spleen (S), kidney (K), white adipose (W), and tail (T) in ob/ob-PPARγ(fl/fl)AlbCre⁺ or ob/ob-PPARγ(fl/fl)AlbCre^– mice. Fragments hybridizing with 3′ probe from the wild-type, floxed, and deleted alleles migrate at approximately 14, 10, and 8 kb, respectively. (c) RNase protection analysis of PPARγ mRNA in OB/OB- or ob/ob-PPARγ(fl/fl)AlbCre mouse livers. Total RNA from three separate mouse livers in each genotype were hybridized with riboprobes for β-actin and PPARγ. The products were then separated on a 5.0% polyacrylamide gel. The size of the protected mRNA fragments for PPARγ and β-actin is as follows; wild-type PPARγ, 195 nt; null PPARγ, 165 nt; and β-actin; 250 nt.