Characteristics of DENV4/2 ED panel and parental strains. (A) Growth kinetics of ED panel on Vero 81 (mammalian) and C6/36 (insect) cells at multiplicity of infection of 0.001 in triplicate. DENV4/2-EDI had significantly lower kinetics compared to other viruses on both Vero 81 and C6/36; Ps < 0.05. All viruses had significantly different growth kinetics on Vero 81 and C6/36 cells, Ps < 0.05. (B) Western blot of virions against envelope (53 kDa) and prM (21 kDa) proteins from stocks grown on Vero 81 and Vero furin-overexpressing (VF) cells, conducted in triplicate. Each representative image consists of Vero 81 and VF stocks of parental strains DENV2 (lanes 1–2), DENV4 (lanes 3–4), and each ED chimera (lanes 5–6), from left to right, DENV4/2-EDI, EDII, and EDIII. (C) Average genome copy number per focus-forming unit (ffu) of mature stocks with RNAse treatment, conducted in duplicate.