Mutagenesis of DNAJC14.

<p>(A) A schematic of DNAJC14 is shown with the truncation mutants indicated below. Arrows above the schematic indicate the location of point mutants engineered in the NT5 truncation mutant backbone. Each mutant contained a C-terminal myc epitope tag, indicated in green. (B and C) SW13 cells were transduced with lentivirus expressing the indicated mutants and 2 d later were challenged with YFV (moi = 5). Cells and media were harvested 1 d later. V, control V1 lentivirus; FL, lentivirus expressing full-length hDNAJC14; WT, the NT5 truncation mutant without any point mutations. (B) Western blot analysis was performed on equal volumes of the cell extracts using anti-myc antibody to detect the tagged DNAJC14 or mutant protein. Migration of size standards (in kDa) is indicated to the left. (C) YFV present in the medium was enumerated by plaque assay. Data represent mean values obtained from triplicate wells; error bars indicate the standard deviation. Pfu, plaque forming units.