Fig. 6

REV-ERB inhibits DENV and ZIKV replication. a Overexpressing REV-ERBα inhibits DENV and ZIKV replication. Huh-7 cells harbouring sub-genomic DENV RNA or ZIKV-Nanoluc were transfected with REV-ERBα expression plasmid or Ctrl empty vector (2 µg). DENV or ZIKV replication was assessed by measuring luciferase activity 48 h later and data expressed relative to untreated Ctrl cells (mean ± S.E.M., n = 4, Mann–Whitney statistical test). REV-ERB expression was confirmed by western blotting. b REV-ERB agonist SR9009 inhibits DENV and ZIKV infection. Huh-7 cells were infected with DENV2 16681 at a MOI of 0.1 and treated with SR9009 for 48 h (n = 3). Extracellular viral RNA and the infectious titre of secreted virus was quantified and the data expressed relative to untreated Ctrl cells, where the mean titre of virus secreted from untreated cells was 1.6 × 10⁵ pfu/ml. Huh-7 cells were infected with ZIKV MP1751 at an MOI of 0.1 and treated with SR9009 for 48 h. Extracellular RNA and infectious titre was quantified and the data expressed relative to untreated cells, where the mean titre of virus secreted from untreated cells was 2.9 × 10⁶ pfu/ml. c A role for SCD in REV-ERB agonist inhibition of DENV and ZIKV replication. DENV replicon cells or ZIKV-Nanoluc infected cells were transfected with CRISPRs targeting exons 2 and 3 of SCD or a scrambled guide RNA and 24 h later treated with SR9009. SCD expression was measured by western blotting and the dose of REV-ERB agonist required to inhibit DENV or ZIKV replication by 50% (IC₅₀) in WT or KD cells determined (mean ± S.E.M., n = 6, Mann–Whitney statistical test)