(A) Sequence of cHP from different strains of DENV2. *, American genotype; †, Malaysia/Indian Subcontinent genotype; ‡, Southeast Asian genotype. R, A or G; Y, C or U. (B) Schematic diagram of pD2/IC variants containing mutations to 3 (LOOPmut3) or 5 (LOOPmut5) of the 7 cHP loop nucleotides. (C) Kinetic analysis of viral replication in D2/IC-, D2/IC-LOOPmut3- and D2/IC-LOOPmut5-transfected BHK cell monolayers. Supernatants were harvested at 24, 48, 72, 96 and 120 hours post-transfection, and viral replication was assessed by plaque assay. PFU are normalized for transfection efficiency. Data are averages of at least 3 experiments. Error bars indicate SD. (D) Schematic of pDRrep-HPhet. (E) Early and late translation events in DRrep-HPhet-transfected cells. Transfected cells were assayed for Rluc activity at 2, 18, 24, 48 and 72 hours post-transfection. RLU are normalized for transfection efficiency as in 5C. Data are averages of 3 experiments and error bars indicate SD. RLU, relative luciferase units. ΔG values are in kcal/mol.