Figure 4. BMI1 activated P- glycoprotein via suppression of miR-3682-3p. (A) qRT-PCR detection of 12 human ABC-transporters, which are associated with drug transport, in T24/DDP&GEM cells upon BMI1 knockdown. (B) Western blot detection of BMI1 and P-GP in T24/DDP&GEM and T24/DDP&GEM-sgBMI1 cells. (C, D) Western blot analysis of BMI1 and P-GP in overexpressing, downregulating BMI1 or vector control cells (T24 and BIU-87). (E) A Venn diagram showing the overlap of candidate miRNAs that were predicted by miRWalk2.0 to potentially bind to the ABCB1 3'-UTR. (F) Heatmap of 6 candidate miRNAs expression in miRNA microarray assay analysis between T24/DDP&GEM and T24/DDP&GEM-sgBMI1 cells. (G) Quantification analysis of miR-3682-3p expression by q-RT-PCR in T24/DDP&GEM and T24/DDP&GEM-sgBMI1 cells. (H) Detection of miR-3682-3p expression by q-RT-PCR in overexpressing, downregulating or vector control cells (T24 and BIU-87). (I) Results of luciferase reporter assay in HEK293T cells with co-transfection of ABCB1 3'-UTR vector (H9688) or mutant control vector (H9689). *P < 0.05. **P < 0.01. ***P < 0.001. DDP: cisplatin; GEM: gemcitabine; P-GP: P-glycoprotein.