Fig. 5: Super-enhancer drives the expression of TCOF1 in TNBC cells.

a Schematic of the putative super-enhancer of TCOF1, indicated by H3K27ac ChIP-seq analysis. e1 and e2 represent constituent enhancers of the super-enhancer predicted by DNase-I hypersensitive seq analysis in MDA-MB-231 cells. b H3K27ac ChIP-qPCR using primers amplifying e1 of TCOF1-associated super-enhancer. Data represent mean ± SEM of three independent experiments. *p < 0.05. c BRD4 ChIP-qPCR using primers amplifying e1 of TCOF1-associated super-enhancer. Data represent mean ± SEM of three independent experiments. *p < 0.05. d Enhancer activity of e1 in HCC1806 and MCF-10-DCIS cells measured by dual-luciferase reporter assay. Data represent mean ± SEM of three independent experiments. **p < 0.01. e Crispr/cas9-mediated DNA deletion of e1 (del-e1) in HCC1806 and MCF-10-DCIS cells. PCR products were analysed by gel electrophoresis. Experiments were repeated twice independently with similar results. f Immunoblotting showing TCOF1 expression in cells with or without e1 deletion. Experiments were repeated twice independently with similar results. g Spheroid growth of HCC1806 and MCF-10-DCIS cells upon e1 deletion. Data represent mean ± SEM of three independent experiments. *p < 0.05; **p < 0.01. h Spheroid growth of HCC1806 with e1 deletion rescued by TCOF1 overexpression. **p < 0.01; ***p < 0.001. i HCC1806 and T47D cells treated with JQ1 were subjected to BRD4 ChIP-qPCR and immunoblotting analysis. Data represent mean ± SEM of three independent experiments. *p < 0.05. p values were calculated by two-sided Student’s t test in b–d, g–i.