FIG 5.
KLF4 is dispensable for BAT development and function. Klf4f/f mice were crossed with Klf4f/+; Myf5-Cre mice to obtain Klf4f/f; Myf5-Cre (conditional KO [cKO]) mice and littermate control (Klf4f/f [f/f]) mice. (A to E) KLF4 is dispensable for brown adipose tissue (BAT) development. Six-week-old cKO and f/f mice were characterized. (A) Genotyping results. The expected ratios of the four genotypes are 1:1:1:1. (B) Body weight of f/f (n = 8) or cKO (n = 13) male mice. (C) Confirmation of Klf4 deletion in BAT by qPCR analysis of genomic DNA. ***, P < 0.001. (D) Pictures of BATs isolated from f/f and cKO mice (n = 3 per group). (E) RNA was extracted from BAT of f/f (n = 8) or cKO (n = 13) mice for qRT-PCR analysis of adipogenesis markers Pparγ, Cebpα, and Fabp4 and BAT markers Prdm16 and Ucp1. Data are presented as means ± SEM. n.s., no significance. (F, G) Cold tolerance test. cKO mice and their littermate controls (n = 5 per group) were acutely exposed to 4°C for 5 h. (F) Body temperatures were measured every hour. (G) qRT-PCR analysis of genes involved in thermogenesis in BAT. RT, room temperature. Data are presented as means ± SEM. (H to K) Characterization of E18.5 embryos. (H) Representative pictures of E18.5 embryos. (I) Confirmation of Klf4 deletion in E18.5 BAT by qPCR analysis of genomic DNA. (J) E18.5 embryos were sagittally sectioned along the midline. The sections of the interscapular area were stained with hematoxylin and eosin (H&E). B, BAT. (K) RNAs were extracted from E18.5 BAT of f/f (n = 7) or cKO (n = 9) embryos for qRT-PCR analysis.