Extended Data Fig. 1: IFI16 induction by IAV is dependent on viral replication.
a–c, IFI16 expression in a, THP-1, b, A549, and c, HEK293 was quantified by RT-qPCR. (d) p204 expression in lung tissues from PR8 virus-infected WT mice was determine. e, IFI16 expression in 1.0 MOI of PR8 virus-infected THP-1 cells was detected. f, IFI16 mRNA expression in 2 MOI of UV-inactivated and live PR8 virus-infected A549 cells was determined. g, IFI16 protein expression in 2 MOI of UV-inactivated PR8 virus-infected A549 cells was determined. h, IFI16 expression in the nuclear and cytoplasmic fractions of PR8 virus-infected A549 cells were determined. β-actin and H3 were used as purity markers for cytoplasmic and nuclear fractions, respectively. i, IFI16 expression in A549 cells transfected with poly(I:C) for 18 h was determined by Western blotting. j, IFI16 expression in A549 cells treated with IFN-γ for 18 h was determined. (k) Intracellular localization of IFI16 in A549 cells treated with poly(I:C) or IFN-γ for 12 h was determined. Scale bars, 45 μm and 5 μm (enlarged). l, A549 cells were infected with PR8 virus at 0, 6, 12 and 24 hpi. Cell lysates were then immunoprecipitated with anti-acetylated lysine. Bound proteins were analyzed by immunoblots with anti-IFI16 antibody. m, A549 cells were pre-incubated with C-646 for 2 h, then infected with PR8 virus for 1 h, washed and incubated in complete medium with or without C-646. IFI16 expression and viral NP protein in the nuclear and cytoplasmic fractions of PR8 virus-infected A549 cells at indicated time points were determined by Western blotting. β-Actin and H3 were used as purity markers for cytoplasmic and nuclear fractions, respectively. (a–d) and f, Data presented as means ± SD from three independent experiments. (e) and g–m, Data are representative of three independent experiments. Statistical significance in (a) to (d) and (f) was determined by unpaired two-tailed Student’s t-test.
