Fig. 2

Cis-regulatory regions in the P3 and P2 promoter of UGT2B17 alt. transcripts. (A) Transcription factor (TF) binding sites predicted by JASPAR [22] in the P3 and P2 promoter regions. STAT3: Signal transducer and activator of transcription 3; NF-κB1: nuclear factor kappa B subunit 1 (p105); PAX5: paired box 5; ESR2: estrogen receptor 2; RORA/B: RAR related orphan receptor A/B; RELA: nuclear factor NF-kappa-B p65; SPI1: transcription factor PU.1; IRF1: Interferon regulatory factor 1; AR: androgen receptor; CEBP: CCAAT/enhancer binding protein. (B) Sequences of the consensus binding sites for STAT3 (MA0144.2), IRF1 (MA0050.1) and RELA (MA0107.1) are shown along with the corresponding sequence in the UGT2B17 P3 or P2 region. Mutations of TF binding motifs for luciferase assays and EMSA (UGT2B17mut) are shown in red. (C) Luciferase (Luc) gene reporter assays with serial deletions of the P3 promoter region. Luciferase activity relative to the pGL3 empty vector (dashed line) is given. (D) Luc assays for P3 promoter activity without (filled circles) or with (open circles) mutated STAT3 and IRF1 binding sites. (E) Luc assays for P2 promoter activity without (filled circles) or with (open circles) mutagenesis of the RELA binding site. In D-E, the luciferase activity relative to the unmutated sequence is shown. Luciferase assays were conducted four times in triplicates. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. Coordinates are relative to the UGT2B17 translation start site (+ 1)