Fig. 3

NF-κB and STAT3 regulate UGT2B17 expression in leukemic cell models. (A) Immunoblot detection of RELA, STAT3 and IRF1 in the MEC1 and JVM2 cell models. Full-length blots are presented in figure S5, additional file 1. (B) Electrophoretic mobility shift assays (EMSA) with biotin labeled oligonucleotides of IRF1, STAT3, or RELA (NF-κB) binding sequences and nuclear extracts of MEC1 and JVM2. The shift of the biotin labeled probes (lanes 4, 9; 14, 19; 24) is impaired by incubation with an excess unlabeled probe (lanes 2, 7; 12, 17; 22) but not with an excess of unlabeled mutated probe (lanes 3, 8; 13, 18; 23). The presence of the RELA antibody produced a supershifted complex (lanes 25). EMSA experiments were conducted twice. Full-length blots and replicates are presented in Figures S6 and S7, Supplementary file 1. (C) inhibition of UGT2B17 mRNA expression by the NF-κB inhibitor BAY 11-7082 in leukemic cell models. (D) Inhibition of UGT2B17 mRNA expression by the STAT3 inhibitor Stattic in leukemic cell models. (E) Inhibition of UGT2B17 protein expression by BAY 11-7082 and Stattic. Protein expression was measured by enzymatic assays after a 24 h-treatment with inhibitors, as described in the Methods section, and is expressed relative to the activity in untreated cells. Two independent qPCR and enzymatic assay experiments were conducted in triplicates. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001