Fig. 5.

Transcription factors NRF1 is involved in insulin-mediated gene expression changes and chromatin remodeling. a Transcription factor-binding motifs enriched in promoter regions (TSS ± 1 kb) of upregulated genes. b Transcription factor-binding motifs enriched in promoter proximal and c distal H3K9ac peaks, respectively. p values for motif enrichment in target sequences over background are mentioned. d–f NRF1 enrichment at NRF1 motifs in promoters of indicated insulin-upregulated genes and g a negative control region determined by ChIP-qPCR in MDA-MB-231 cells treated with insulin (100 nM) for 3 h or 6 h. IgG and FOXK2 antibody ChIPs serve as negative controls. Bars represent percent input pull-down in untreated (UT) and treated (3 h, 6 h) cells. Values are Mean + SEM from two independent experiments and three technical replicates from each experiment. Statistical significance was calculated using one-way ANOVA, Dunnett’s multiple comparisons test. ****p < 0.0001. h–k Quantification of expression of NRF1 (h) and NRF1 target genes (i–k) after siRNA-mediated NRF1 knockdown with or without insulin treatment for 6 h in MDA-MB-231 cells. Values are Mean + SEM from three independent experiments. Statistical significance was calculated using one-way ANOVA, Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ****p < 0.0001, ns = nonsignificant