Fig. 2: Hyperglycemia establishes active enhancer histone marks on the Nrg1 enhancer.

a Graphic representation of Nrg1 gene and its enhancer. The Nrg1 enhancer spans 0.6 kb and is located on mouse Chromosome 8: 32,200,290–32,199,690. The center of the enhancer is marked “0”, and primer pairs are shown in identical colors. b, c ChIP-qPCR analysis of H3K27ac and H3K4me1 enrichment at the Nrg1 enhancer in Met1 cells treated with low glucose (LG) or high glucose (HG). d qPCR analysis of chromatin accessibility of the Nrg1 enhancer (+200 bp) in Met1, 4T1, and Eo771 cancer cells. e, f mRNA and protein levels of Nrg1 in trichostatin A (TSA)-treated Met1 cells under LG. g–j ChIP-qPCR analysis of time-course binding occupancy of P300, CBP, SETD1A, and HDAC1 at the Nrg1 enhancer in HG-treated Met1 cells. k Western blot analysis of subcellular protein localization of SETD1A and HDAC1 in LG- and HG-treated Met1 cells. l Protein levels of NRG1 in non-treated, shScrb, shP300, shCbp, and shSetd1A Met1 cells after up to 72 h of HG treatment. m Cellular proliferation of HG-treated cancer cells expressing scrambled or Setd1A shRNA. Data represented mean ±SEM of technical replicates in b–e, g–j, and biological replicates in m, and n = 3 independent experiments were performed. Statistical significance was evaluated using two-way ANOVA followed by Bonferroni’s b, c, d, Tukey’s g, h, j, m, and Dunnett’s i post hoc tests, and one-way ANOVA followed by Tukey’s post hoc test e. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001 for column 1 versus other columns and ^#P  < 0.05; ^##P < 0.01; ^###P < 0.001 for column 2 versus column 4, b, c (all ^***P < 0.0001), d (from left to right, ^*P = 0.0413, ^**P = 0.0088, ^***P < 0.0001), e (from left to right, ^**P = 0.0042, ^**P = 0.0026, ^***P < 0.0001), g (from left to right, ^***P < 0.0001, ^*P = 0.0367, ^*P = 0.0189), h (from left to right, ^*P = 0.0326, ^***P = 0.0002, ^**P = 0.0023), i (from left to right, ^***P = 0.0004, ^*P = 0.0226, ^*P = 0.0276, ^**P = 0.0025, ^*P = 0.0239, ^*P = 0.0407, ^*P = 0.0358, ^**P = 0.0025), j (from left to right, ^*P = 0.0313, ^***P < 0.0001, ^*P = 0.0184, ^***P = 0.0006, ^***P = 0.0028, ^*P = 0.0264), m (^*P = 0.0141, ^#P = 0.025). Source data are provided in Source Data file.