Figure 6
Administration of FGF21-mimetic antibody phenocopies cold exposure in obese VAT. HFD-fed male mice received bFKB1 (10 mg/kg i.p.) or vehicle once on day 0 at thermoneutral condition. Tissues were harvested on day 14 after bFKB1 injection for histology and FACS analysis (A–F), and day 8 for gene expression analysis (G–J). (A) H&E staining in HFD-fed SCAT. (B) H&E staining in HFD-fed VAT. (C) Representative plots for M2/M1 macrophage populations. (D) Ratio of the frequencies of ATM2 (CD11c−/CD206+) over ATM1 (CD11c+/CD206−). (E) CD206 (green) and Arg1 (red) IF. (F) Lyve1 (green) and Arg1 (red) IF. DAPI (blue) stains cell nuclei. (G) tSNE analysis of RNA-Seq results from ATM1 and ATM2 from obese VAT. (H) 4-way differential analysis revealed bFKB1-induced gene expression profile in ATM2 mimics cold exposure. Grey dots indicate all the expressed genes. Red dots indicate genes of interest. (I) Expression of genes regulated by bFKB1 in ATM1 or ATM2. Data are shown as mean ± SEM (N = 3 samples, 5 mice pooled as one sample). (J) Heatmap showing the expression of bFKB1-regulated genes in ATM2. The gene set was selected based on a 2-Way comparison between bFKB1-treated samples and control (warm) samples. Color scale corresponds to the relative expression level (log2(RPKM) standardized along each gene). The gene names are provided in Figure S9. *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t-test.
