Fig. 4.

REV-ERB agonists modulate miR-122. a Pri-miR122 displays circadian pattern in synchronised Huh-7 cells. qRT-PCR analysis of pri-miR-122 levels relative to GAPDH in cycling Huh-7 cells (mean ± S.E.M., n = 3). b REV-ERB agonists reduce pri-miR122 levels. qRT-PCR analysis of pri-miR-122 relative to GAPDH in Huh-7 cells treated with REV-ERB agonists (20 μM) for 16 h (mean ± S.E.M., n = 3, Kruskal–Wallis ANOVA with Dunn’s test). c Anti-viral activity of REV-ERB agonists is miR-122 dependent. Huh-7 cells were electroporated with HCV JFH-Luc and miR-122 antagonist (100 pM per 10⁶ cells) or a randomised control, allowed to recover for 4 h and treated with REV-ERB agonists (20 μM) for 24 h. Hepatitis C virus (HCV) replication was assessed by measuring luciferase activity and the data expressed relative to Ctrl untreated cells (Mean ± S.E.M., n = 4, Kruskal–Wallis ANOVA with Dunn’s test). d REV-ERB agonists inhibit HCV independent of direct miR-122 binding. Huh-7 cells were inoculated with WT or miR122 independent HCV (m15) for 6 h, unbound virus removed by washing and the infected cells treated with SR9009 or GSK2667 for 48 h and the frequency of NS5A expressing cells quantified and expressed relative to untreated (Ctrl) cells (mean ± S.E.M., n = 3)