FIG 6.

DDX21 knockdown results in the accumulation of R-loops on HCMV viral late gene. (A) MRC5 cells were transduced with lentivirus vectors respectively expressing shC, shDDX21-1, or shDDX21-3. After 48 h postransduction, the cells were infected with HCMV at an MOI of 0.1. Cells were fixed at 72 hpi and stained with antibodies against S9.6 (marker of R-loops) and DDX21. A bar graph represents the mean intensity of S9.6 fluorescence in the nuclei of individual cells (n = 20). ***, P < 0.001. (B to D) DRIP qPCR assay monitored R loops on viral late gene UL32 (B), viral immediate early gene IE1 (C), and viral early gene UL44 (D) in cells upon siRNA-mediated depletion of DDX21. The 2^–ΔCT value is presented as DRIP (% input): ΔC_T = C_T DRIP – C_T Input; ΔC_T (RNase H) = C_T DRIP (RNase H) – C_T Input (RNase H). *, P < 0.05; ns, not significant.