Fig. 2
miR-29b-1-5p targets NTCP transcripts in human hepatocytes. (A) miR-29b-1-5p overexpression. PHHs were transfected using miR-29b-1-5p mimics (miR-29b-1-5p) and miRNA control mimics (miR-Control). Relative expression levels were measured by real-time quantitative PCR after four days. miR-29b-1-5p expression levels were normalized using RNU6B. (B) Effect of miR-29b-1-5p overexpression on NTCP mRNA levels. miR-29b-1-5p expression was measured four days after transfection using synthetic miRNA mimics. NTCP expression levels were normalized using GAPDH. (C) NTCP 3ʹ-UTR dual luciferase reporter assay. Human hepatoma HepG2 cells were co-transfected with miRNA mimics (miR-29b-1-5p versus miR-Control) and luciferase reporter constructs containing the human NTCP 3ʹ-UTR (WT, wild type) or a mutated sequence (Mut, mutated). The luciferase activities were measured 24 h after transfection, and the ratio of hRluc/hluc+ was determined. The depicted sequences indicate sequence complementarity between miR-29b-1-5p and NTCP 3ʹ-UTR. All histograms represent the mean ± SD. Statistical differences were evaluated with a t-test for miR-29b-1-5p expression measurements (n = 6 per condition) and the luciferase activities (n = 3 per condition) and a one-way analysis of variance (ANOVA) for NTCP expression measurements (n = 6 per condition). *p < 0.05, **p < 0.01. NS, not significant.
