Identification and Characterization of the Host Protein DNAJC14 as a Broadly Active Flavivirus Replication Modulator
Figure 3
DNAJC14 inhibits YFV replication.
(A). YFV replication is inhibited in Round 3 (Rd 3) cells. Rd 3 or naïve cells were infected with YFV (moi = 1) and at the indicated days after infection the amount of virus released into the medium was determined by plaque assay. A single well was used for each cell type, with replacement of the medium at each timepoint. (B). Hamster DNAJC14 inhibits YFV protein expression. SW13 cells were transduced with control lentivirus (V1-GFP) or lentivirus containing the truncated hamster DNAJC14 insert (clone 1-2, 1-3, 1-4 and 1-5) and were infected 2 d later with YFV (moi = 0.5). Cells were analyzed by Western blot for NS3 expression 3 d after infection. Naïve cells serve as a negative control and actin serves as a loading control. (C). Full-length and truncated human DNAJC14 inhibit YFV replication. SW13 cells were mock transduced (Mock) or transduced with the indicated lentiviral vectors and 2 d later were infected with YFV (moi = 0.5) as indicated. Cells were harvested and subjected to Western blot analysis 3 d after infection using antibodies as indicated to the right. V1-FL expresses myc epitope tagged full-length human DNAJC14 while V1-NT1 expresses myc tagged truncated human DNAJC14 (aa 305–702). Naïve cells were left untransduced and uninfected and serve as a negative control. For B and C migration of size standards (in kDa) is indicated.