rs9952980 affects SLC14A2 expression via reduced ESR1 binding by impacting ERE. (A) Snapshots of ESR1 peak intersecting the coordinate of rs9952980. The peak, positioned in an intron of SLC14A2, was found in 11 female patients. (B) ERE at this peak, in reference allele and rSNP format. (C) Predicted score of position weight matrix for WT and rSNP ERE, by SNP2TFBS (Kumar et al. 2017). (D) Using MCF-7 lysate, an immunoprecipitation (IP) was performed with 50-bp biotin-labeled oligos containing the WT or the rs9952980 variant of the ERE, followed by mass spectrometry. (E) ESR1 western blot (WB) of IP by 50-bp biotin-labeled oligos containing the reference allele or the rs9952980 variant of the ERE. (F) Snapshot of STARR-seq normalized signal at the SLC14A2 locus. (G) Luciferase reporter assay in MCF-7 cells stimulated or not by estradiol (E2) for the SLC14A2 locus enhancer activity containing or not the rs995290 variant. Bar plot represents the fold change of luciferase expression over the untreated empty vector condition. (H) TCGA gene expression of SLC14A, which rs9952980 is predicted to affect, by homozygous or heterozygous genotype.
