FIG 6.
Krox20 is dispensable for BAT development. Krox20f/f were crossed with Krox20f/+; Myf5-Cre to obtain Krox20f/f; Myf5-Cre (conditional KO [cKO]) and littermate control (Krox20f/f [f/f]) mice. (A to D) Krox20 is dispensable for BAT development. Eight-week-old cKO and f/f mice were characterized. (A) Genotyping results. The expected ratios of the four genotypes are 4:1:1:4. (B) Isolated adipose tissues. iWAT, interscapular WAT; epi-WAT, epididymal WAT; ing-WAT, inguinal WAT; rWAT, retroperitoneal WAT. (C) Confirmation of Krox20 deletion in BAT by qPCR analysis of genomic DNA. (D) RNA was extracted from BAT of f/f and cKO mice for qRT-PCR analysis of adipogenesis markers Pparγ, Cebpα, and Fabp4 and BAT markers Prdm16 and Ucp1. (E to G) Characterization of E18.5 embryos. (E) Representative pictures of E18.5 embryos. (F) Krox20 deletion in BAT was confirmed by qRT-PCR of genomic DNA. (G) H&E staining of the interscapular area. B, BAT. (H) Krox20 mRNA is largely absent in BAT. The mRNA levels of early (GR, Cebpβ, Cebpδ, Klf4, and Krox20) or late (Pparγ, Cebpα, Fabp4, Prdm16, and Ucp1) adipogenesis genes in BATs isolated from adult (8-week-old) mice or embryos (E18.5) were determined by RNA-Seq. RPKM values from RNA-Seq indicate gene expression levels (log10 scale). ***, P < 0.001; n.s., no significance.