Fig. 5. Obesity repositions BMAL1 binding genome-wide in human adipocyte precursors.

A Average plot of BMAL1 ChIP-seq reads on the target gene promoters near TSS in OAPs from non-obese (n = 3; blue) and obese (Class III) patients (n = 3; red). B Top known HOMER motifs enriched at BMAL1-binding sites from ChIP-seq analysis in OAPs from non-obese and obese patients. The highest ranking motif for each patient is shown and corresponds to the canonical E-box motifs. Significant motif enrichment was determined using a cumulative binomial statistical test. C–F University of California at Santa Cruz (UCSC) genome browser images of BMAL1 ChIP-seq tracks at clock repressors and related histograms. UCSC genome browser images of BMAL1 ChIP-seq tracks at PER2 (C) and REV-ERBα (E). Normalized tag counts are indicated on the Y-axis and maximum track height is the same for all samples (n = 3 subjects/group). The orientation for each gene is indicated below each browser track. D, F Respective histograms depicting the average peak value per condition are also represented. Data are means ± SEM, *p < 0.05, by unpaired two-tailed t-test. G VENN diagram depicting the number of BMAL1 peaks and overlap in non-obese and obese OAPs (Raw data in OAPs from 3 patients/ group are provided in Source Data). H Average plot of BMAL1 ChIP-seq reads depicting their distance to NF-κB consensus motifs on target genes. NF-κB sites were obtained from UCSC Genome Browser’s Factorbook data, which were lifted over from hg38. The normalized signal files for each group (average n = 3 subjects/group) were merged to show that BMAL1 binds in close proximity to NF-κB consensus motifs. See also Supplementary Fig. 4.