Figure 3.

Hfe deficiency enhances type I IFNs induction in response to H7N9 virus infection. (A) The mRNA transcription of the genes Ifnb1, Il6 and Tnf in the lungs of H7N9 virus-infected WT and hfe^−/- mice were analyzed by qRT-PCR at 2 dpi. The concentrations of IFNB1, IL6 and TNF were evaluated in the serum and BAL at 2 dpi (n = 5). (B and C) The heatmaps showed 30 genes that were differentially expressed in the BMDMs of H7N9 virus-infected WT and hfe^−/- mice and that were involved in the innate response (B) and inflammation (C). (D and E) qRT-PCR and ELISA analysis of the mRNA and expression of the genes Ifnb1, Il6 and Tnf in BMDMs infected with H7N9 virus at the indicated time points. (F) TCID₅₀ was used to measure the H7N9 virus titers in the supernatants of WT and hfe^−/- BMDMs infected with H7N9 virus. (G and H) qRT-PCR and ELISA analyzed the mRNA and expression of the genes Ifnb1, Il6 and Tnf in BMDMs of WT or hfe^−/- mice transfected with poly (I:C) (1 μg/ml) or HSV60 (1 μg/ml) for 4 h. (I) Immunoblot analysis of the indicated proteins in lysates of WT and hfe^−/- BMDMs infected with VSV or H7N9 virus. (J) Immunoblot analysis of the indicated proteins in lysates of WT and hfe^−/- BMDMs transfected with poly (I:C) (1 μg/ml) or HSV60 (1 μg/ml) for 4 h. All qRT-PCR data are normalized to the Actb gene and used the control group as a reference. Data are representative of at least three independent experiments, and differences between the experimental and control groups are determined by 2-way ANOVA (*p < 0.05, **p < 0.01, and ***p < 0.001)