Fig. 3: Adipocyte Cldn5 ablation represses thermogenesis and energy expenditure.
From: Adipocyte CLDN5 promotes thermogenesis and energy expenditure through regulation of IL10 expression
a Volcano plot of RNA-seq data in BAT showing gene expression differences between Cldn5flox/flox and Cldn5flox/flox; Fabp4-Cre mice, analyzed by the DESeq2 R package (1.16.1). b Top enriched biological processes of RNA-seq data in BAT determined by Gene Ontology analysis using clusterProfiler 3.8.1. c–g qRT-PCR analysis of indicated genes related to thermogenesis and oxidative phosphorylation (OXPHOS) (c, n = 5), Western blot analysis of PGC1α, UCP1 (d, n = 3) and OXPHOS proteins (e, n = 3), mitochondrial DNA (mtDNA) copy number (f, n = 8), and transmission electron microscopy (g) of BAT from Cldn5flox/flox and Cldn5flox/flox; Fabp4-Cre mice. h–k Energy expenditure (h), food intake (i), physical activity (j), and respiratory exchange rate (RER, k) of Cldn5flox/flox (n = 4) and Cldn5flox/flox; Fabp4-Cre mice (n = 5). l, m Rectal temperature (l), infrared thermal images and quantification of BAT interscapular temperature (m) of Cldn5flox/flox (n = 14) and Cldn5flox/flox; Fabp4-Cre mice (n = 13) challenged at 4 °C for 4 h. n–p Rectal temperature (n, n = 7), infrared thermal images and quantification of BAT interscapular temperature (o, n = 7), and free fatty acid (FFA) levels (p, Cldn5flox/flox, n = 5; Cldn5flox/flox; Fabp4-Cre, n = 6) in Cldn5flox/flox and Cldn5flox/flox; Fabp4-Cre mice kept at 4 °C for 7 days. q, r qRT-PCR analysis (q) and Western blot analysis (r, n = 3) of PGC1α and UCP1 in Cldn5flox/flox and Cldn5flox/flox; Fabp4-Cre mice housed at RT or 4 °C for 7 days. q, BAT: Pgc1α (n = 7), Ucp1 (n = 6); iWAT: Pgc1α (n = 5), Ucp1 (n = 7). s, t qRT-PCR analysis (s, n = 5) and Western blot analysis (t, n = 3) of PGC1α and UCP1 in Cldn5flox/flox and Cldn5flox/flox; Fabp4-Cre mice treated with saline/CL-316,243. u, v HE staining with droplet size quantification in the mice shown in panels n–r (u) and s, t (v), n = 5. Scale bar, 5 μm (g) or 50 μm (u, v). Data represent the mean ± SEM. Statistical analyses were performed using Mann-Whitney test (Atp5e in c, o), unpaired t test (other genes in c; d–f, l–n), two-way ANOVA with Tukey’s post-hoc test (q–v), ANCOVA (h, i), or ANOVA (j, k). All tests were two-sided. N, nucleus; LD, lipid droplet. Source data are provided as a Source Data file.
