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. 2024 Aug 13;15:6964. doi: 10.1038/s41467-024-51192-1

Fig. 3. SARS-CoV-2 N protein induces DNA damage by downregulating Dicer, XPO5, SRSF3, and hnRNPA3 expression.

Fig. 3

See also Supplementary Figs. 3, 4. a–c DNA damage in A549-Ctrl or A549-N cells was determined using immunoblotting analysis of the phosphorylation levels of ATM, ATR, Chk1, Chk2, and H2AX (a), comet assay (b), and immunofluorescence with anti-γ-H2AX antibody (c). d, e A549-hACE2 cells infected with SARS-CoV-2 or mock were subjected to immunofluorescence with anti-γ-H2AX antibody (d) and immunoblotting with indicated antibodies (e). A549 cells were transfected with the indicated siRNAs and subjected to comet assay (f) and immunofluorescence with anti-γ-H2AX antibody (g). h A549-Ctrl and A549-N cells transfected with a control plasmid or plasmids overexpressing Dicer, XPO5, SRSF3, hnRNPA3, or RNase H1 were subjected to comet assay. i Detection of R-loop in A549-Ctrl or A549-N cells using immunofluorescence with the S9.6 anti-DNA-RNA hybrid antibody. j Detection of R-loop in A549-hACE2 cells infected with SARS-CoV-2 or mock using immunofluorescence with the S9.6 anti-DNA-RNA hybrid antibody. k Detection of R-loop in A549-Ctrl or A549-N cells transfected with a plasmid expressing RNase H1 or control plasmid using immunofluorescence with the S9.6 anti-DNA-RNA hybrid antibody. Data in (b–k) are expressed as mean ± SD (two-tailed unpaired Student’s t-test). n = 100 (b, f, h) or 200 (c, d, g, i–k) cells from three biological replicates. **p < 0.01; ns not significant (p > 0.05; two-tailed unpaired Student’s t-test). The numbers below the blots (a, e) represent the relative densitometric quantification of the bands normalized to corresponding GAPDH bands; the mean values of three independent experiments are shown. Ctrl control plasmid, N N protein, siNC negative control siRNA, si-all cells transfected with siDicer, siXPO5, siSRSF3, and sihnRNPA3 together, OE-all cells transfected with Dicer-, XPO5-, SRSF3-, and hnRNPA3-expressing plasmids together. Source data and exact p values are provided in the Source Data file.