Figure 1.

Influenza Virus Triggers Oxidized DNA Release from Macrophages

(A) J774A.1 macrophages were subjected to digitonin fractionation as described in Methods and pellets (P) or cytosolic extracts (C) were analyzed by western blotting using the indicated antibodies.

(B–E) J774A.1 macrophages were infected with PR8 virus. Pure cytosolic extracts and supernatants were collected at indicated time points. Relative levels of mtDNA (B and D) or nuclear DNA (C and E) in the cytosol and supernatants were assessed by quantitative PCR.

(F) J774A.1 macrophages were infected with PR8 virus. At 24 h post infection, cells were stained with anti-dsDNA (AC-30-10) and anti-Tom20 antibodies and analyzed by confocal microscopy. Scale bars, 10 μm.

(G) J774A.1 macrophages were infected with PR8 virus. Pure cytosolic extracts were collected at indicated time points. Oxidized DNA in the cytosol was detected by dot blot analysis using anti-8OH-dG antibody. These data are from three independent experiments (B–E; mean ± SEM).

∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 versus mock-infected cells (one-way ANOVA and Tukey's test). See also Figure S1.