DNAJC14 inhibits multiple members of the Flaviviridae family.

SW13 (A, B) or Huh7.5 (C–E) cells were seeded in equal numbers into plates and transduced with V1 vector expressing GFP (GFP, filled circles), V1 vector expressing full-length human DNAJC14 (FL, open circles) or V1 vector expressing truncated (aa 305–702) human DNAJC14 (NT1, triangles). After 2 days, the cells were challenged (moi = 5) with YFV 17D (A), YFV Asibi (B), Kunjin virus (C), Langat virus (D) or were challenged (moi = 0.1) with HCV Jc1FLAG2/p7-nsGluc2A. The medium (A–D) or cells (E) were harvested at the indicated times for quantification of virus replication. For A and B, a single separate well was utilized for each time point, and virion production was enumerated by plaque assay. In both cases, there were fewer than 100 plaque-forming units at 12 h post infection. The dashed line indicates the sensitivity of the plaque assay. Pfu, plaque forming units. For C and D, duplicate wells were infected and the medium was harvested and replaced at each timepoint. Virion production since the prior time point was enumerated by focus forming assay as described in Materials and Methods. Data points represent the mean titer; error bars indicate the range. Similar results were obtained in an independent experiment for both Kunjin and Langat viruses. FFU, focus forming units. For E, cells were harvested at the indicated times after infection for measurement of luciferase activity as described in Materials and Methods. Data points represent mean values obtained from triplicate wells; error bars indicate the standard deviation. RLU, relative light units.