Fig. 5: Regulation of enhancer activity by EBF2 and CBP is conserved between mice and humans. | Communications Biology

Fig. 5: Regulation of enhancer activity by EBF2 and CBP is conserved between mice and humans.

From: Identification of a distal enhancer of Ucp1 essential for thermogenesis and mitochondrial function in brown fat

Fig. 5

a Venn diagram showing the transcription factors of Ucp1-En4 and control group identified using DNA pulldown assay in two replicates of iBAT. b ChIP-seq profiles showing binding of different transcription factors to Ucp1-En4 region. c ChIP-qPCR analyses showing the binding of EBF2 and CBP to Ucp1-En4 region in control and EBF2-overexpressing brown adipocytes (n = 3). d ChIP-qPCR analyses showing enrichment of H3K27ac on Ucp1-En4 region in control and EBF2-overexpressing brown adipocytes (n = 3). e qRT-PCR analysis of Ucp1 mRNA expression in control and EBF2-overexpressing brown adipocytes (n = 5). f H293T cells were co-transfected with Ucp1-En4 construct with EBF2 or CBP either together or individually (n = 6). g ChIP-seq (including H3K27ac, CBP, and MED1) and DHS-seq profiles defined putative active enhancers of UCP1 in beige adipocytes differentiated from human multipotent adipose-derived stem (hMADS) cells. Putative active enhancer is represented by an orange rectangle. h TF binding site motif analysis on Ucp1-En4 and UCP1-hEn1 sequences using the JASPAR database (https://jaspar.genereg.net/). i H293T cells were co-transfected with UCP1-hEn1 construct with human EBF2 or CBP either together or individually (n = 6). Data are presented as mean ± SEM. P values were calculated by two-tailed unpaired Student’s t-tests.

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