CTCF regulates glucose metabolism and stress response related gene expression in murine islets. (a) Relative CTCF mRNA expression in Min6 cells treated with indicated reagents for 12 h. Data represent mean ± SEM; n = 3–4 biological replicates; expression levels were normalized to their respective controls; *, P < 0.05; **, P < 0.005; ***, P < 0.001; one-way ANOVA; Veh, Vehicle. (b) Immunoblots of total cell lysates from Min6 cells treated with PA and TNFα. (c and d) Heatmap showing scaled, normalized read-counts (as Z-Score) of significantly reversible CTCF bound peaks in islets between HFD vs. Chow and HFD-Chow vs. HFD comparison and GO analysis of genes annotated to these peaks. Most significant and nonredundant biological processes with respective gene number and P value are shown. (e) Overlapping analysis of significantly altered genes in islets isolated from BKS-db/db mice treatment with AAV-CTCF or saline via pancreatic ductal infusion and reversible genes by dietary intervention. Heatmap representation of significantly altered genes between AAV-CTCF and saline treatments (left, normalized read-counts were shown as Z-Score), Venn plot between two datasets (middle), and heatmap representation of overlapped genes with similar regulatory trends (right) are shown. (f) Representative RNA-Seq and ATAC-Seq browser tracks displaying dietary intervention and CTCF overexpression regulated gene loci including Hspa1a, Hspa1b, and Slc2a2. CTCF CUT&Tag tracks and ENCODE annotated CREs are also displayed. (g) Schematic representation of the working model presented in this paper. Data in a and b are representative of at least two independent experiments. Source data are available for this figure: SourceData F7.