The H3K27ac region containing rs55958994 (chr12: 53300245 to 53303204, UCSC.hg19) was amplified from 22Rv1 genomic DNA and cloned into the pGL3-Promoter vector (E1761, Promega) upstream of SV40 promoter; the region for analysis was selected on the basis of the H3K27ac ChIP-seq data from 22Rv1 cells. The introduction of the T versus C allele at rs55958994 was obtained by site-directed mutagenesis. Reporter plasmids and the pRL-TK Renilla luciferase control vector (E2241, Promega) were cotransfected into 22Rv1, C4-2B, or LNCaP cells using Lipofectamine 3000 reagent (Invitrogen). Cells were harvested for luciferase assays, and luciferase activity was determined using the Dual-Luciferase Reporter Assay System (E1910, Promega); the luminescent signals from experimental samples were normalized to controls.

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