Journal: PLOS Pathogens

Article Title: Fructose-1,6-diphosphate inhibits viral replication by promoting the lysosomal degradation of HMGB1 and blocking the binding of HMGB1 to the viral genome

doi: 10.1371/journal.ppat.1012782

Figure Lengend Snippet: (A and B) RAW264.7 cells (A) and BMDMs (B) were treated with the indicated concentration of FBP for 12 h and then infected with VSV (MOI, 0.1) for 10 h. The cells were collected, and the expression of Ifnb1 mRNA was assessed by qPCR. (C) HLCZ01 cells were infected with HCV (MOI, 0.01) for 72 h and treated with 1 mM FBP for 12 h. The cells were collected, and the mRNA levels of IFNB1 were assessed by qPCR. (D) BMDMs were treated with 1 mM FBP for 12 h and then infected with HSV-1 (MOI, 0.1) for 10 h. The cells were collected, and the mRNA levels of Ifnb1 were assessed by qPCR. (E and F) RAW264.7 cells were treated with 1 mM FBP for 12 h and transfected with HT-DNA or 100 bp dsDNA for 8 h (E) or poly(I:C) (20 μg/mL) for 12 h (F), and the mRNA levels of Ifnb1 were assessed by qPCR. (G and H) BMDMs from IFNAR1-WT and IFNAR1-KO mice were treated with 5 mM FBP for 12 h and then infected with VSV (MOI, 0.1) (G) or HSV-1 (MOI, 0.1) (H) for the indicated times. Western blot analysis of the VSV-G protein and qPCR analysis of HSV-1 gDNA levels were performed. (I) Huh7.5 cells were infected with HCV at the indicated MOI for 72 h and treated with 5 mM FBP for 12 h. The cells were collected, and the RNA levels of HCV were assessed by qPCR. (J-L) C57BL/6 mice were pretreated with FBP (500 mg/kg) or PBS (every 12 h for two days) and then intraperitoneally injected with VSV (J and K) or HSV-1 (L) (2×10 8 PFU per mouse). FBP or PBS was injected at the same frequency during viral infection. Mice were sacrificed at 24 h (J and K) or 48 h (L) after infection to analyze serum IFN-β concentrations by ELISA (J) or the mRNA levels of Ifnb1 in the liver (K) and brain (L) by qPCR. Data are presented as the mean ± SEM. NS, not significant, * p < 0.05; ** p < 0.01; *** p < 0.001. one-way ANOVA. In (H), statistical analyses were performed with one-way ANOVA. In (A-F), (I) and (J-L), statistical analyses were performed with two-tailed Student’s t test.

Article Snippet: The concentrations of IFN-β in the serum were measured using a mouse IFN-β ELISA kit (R&D Systems), according to the manufacturer’s instructions.

Techniques: Concentration Assay, Infection, Expressing, Transfection, Western Blot, Injection, Enzyme-linked Immunosorbent Assay, Two Tailed Test