FIG 3.

shRNA knockdown of DDX21 inhibits HCMV replication. (A) Knockdown efficiency of shRNAs targeting DDX21 genes. Cells were transduced with lentivirus vectors expressing shC, shDDX21-1, or shDDX21-3. Cell lysates were collected at 48 h postransduction, and the protein levels of DDX21 were determined by Western blot analysis. (B) Growth analysis of HCMV infection in cells expressing shC, shDDX21-1, or shDDX21-3. The cells were infected at an MOI of 1, the cell-free viruses in the supernatant were collected at the indicated times after infection, and the titer was determined using a TCID₅₀ assay. Statistical analysis showed that DDX21 knockdown inhibited HCMV replication. ***, P < 0.001. (C) Growth analysis of HCMV infection in cells expressing shC, shDDX21-1, or shDDX21-3 at an MOI of 0.1. The experimental procedures used were similar to those described for panel B. A statistical analysis showed that DDX21 knockdown inhibited HCMV replication. ***, P < 0.001. (D) Knockdown efficiency of these two shRNAs in cells expressing shC, shDDX21-1, or shDDX21-3 at 3 and 6 dpi. The cells were infected at an MOI of 1, the cell lysates were collected at indicated time points, and the protein levels of DDX21 were detected by Western blot analysis. (E) Detection of knockdown efficiency of coexpression of these two shRNAs. Cells were transduced with lentivirus vectors expressing shC, shDDX21-1, shDDX21-3, or both shDDX21-1 and shDDX21-3. Cell lysates were collected at 48 h postransduction, and the protein levels of DDX21 were determined by Western blot analysis.