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. 2022 Dec 5;119(50):e2214988119. doi: 10.1073/pnas.2214988119

Fig. 3.

Fig. 3.

Changes in accessibility and interactions at two CREs in the Fto locus correlate with transmission of obesity. (A) Organization of the region surrounding the Fto gene and its relationship to Fig. 3 B and C. (B) Persistence of an ATAC-seq accessible site (Site 1) located in intron 8 of the Fto gene in sperm of BPA-F1 through BPA-F5 males. This site contains binding motifs for CTCF and FOXA1, and it is also present in sperm of the male progeny from the IVF experiment. Other sites in the region are present in CTL and BPA-F6 but they increase in ATAC-seq signal in sperm of BPA-F1 through BPA-F5 (Top). ChIP-seq experiments confirm the presence of CTCF at site 1 in sperm of BPA-F3 but not control males (Bottom). (C) Differences in chromatin accessibility at the Fto distal CRE between sperm from BPA-F6 and unexposed controls versus BPA F1-F5 males. Differentially accessible sites contain motifs for CTCF and AR. (D) Significant differences in interaction frequency between the Fto proximal CRE and other sites in the genome using anchors identified using significant interactions determined by Fit-Hi-C and all Hi-C interactions in sperm of CTL and BPA-F3 mice. The 31 significant Fit-Hi-C interactions between the Fto proximal CRE and other sites were analyzed using a bullseye transformed metaplot that depicts distance normalized Hi-C signal in each dataset (P-value = 0.019; unpaired two sample t test). Scores represent average distance-normalized signal. (E) Differential significant Hi-C interactions between the Fto proximal CRE and adjacent genomic sites in sperm of BPA-F3 and CTL mice. The Y-axis represents the difference between the distance-normalized Hi-C signal in sperm of BPA-F3 mice and control. (F) Differences in interaction frequency between the Fto distal CRE and other sites in the genome using significant interactions determined by Fit-Hi-C and all Hi-C interactions in sperm of CTL and BPA-F3 mice. Ten significant Fit-Hi-C interactions were identified, and the bullseye metaplot score represents average distance-normalized signal in each dataset (P-value = 0.298; unpaired two sample t test). (G) The Fto distal CRE preferentially interacts with the Fto and Irx5genes, including the proximal CRE, in sperm of BPA-F3 versus CTL. The Y-axis represents the difference between the distance-normalized Hi-C signal in sperm of BPA-F3 mice and control (SI Appendix, Fig. S3).