Fig. 2: Direct role for REV-ERBα to bind and regulate NTCP.

a HepaRG cells were co-transfected with NTCP promoter luciferase reporter (−1kb) and control or Halo-tagged Rev-Erbα expression plasmid. NTCP promoter activity was determined 48 h later by quantifying luciferase activity and data expressed relative to control (mean ± SEM, n = 5, Mann–Whitney test, Two-sided). b dHepaRG cells were treated with SR9009 (20 µM) for 24 h and NTCP RNA and protein measured along with the housekeeping β-actin by qRT-PCR or western blotting (mean ± SEM, n = 5, Mann–Whitney test, Two-sided). c dHepaRG cells were treated with SR9009 (20 µM) for 24 h; [3H]-taurocholic acid was supplied with media for 10 min; intracellular radioactivity was measured by liquid scintillation counting and data expressed relative to control (mean ± SEM, n = 3, One-way ANOVA with multiple comparisons, Two-sided). Coordinates of RORE or E-box motifs in NTCP promoter. Chromatin extracts from HepaRG cells were immunoprecipitated using antibodies specific for REV-ERBα (d), BMAL1 (e) or rabbit IgG as a negative control. PCR for NTCP promoter regions using primers targeting defined RORE or E-box motifs and control host genes Bmal1 or Per1 was performed and %IP data presented relative to a rabbit IgG control shown as the dotted line (mean ± SEM, n = 4, Mann–Whitney test, Two-sided). *p < 0.05, **p < 0.01. Data are provided in the accompanying Source Data file.