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Image Search Results
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) A simplified model describing the metabolic processes that regulate FBP production and consumption. FBP: Fructose-1,6-diphosphate, PFK1: Fructose-6-phosphate kinase 1, FBP1: Fructose-1,6-bisphosphatase 1, PEP: Phosphoenolpyruvate, Ox Phos: Oxidative Phosphorylation. (B-P) 293T cells were transfected with various constructs including siControl, siFBP1 (B-D), an empty vector (EV), PFKP-overexpressing plasmid (E-G), FBP1-overexpressing plasmid (H-J), siPFKP (K-M), FBP1 enzyme activity deletion mutant (FBP1-G260R), nuclear localization mutant (FBP1-NLS), and nuclear localization with activity deletion mutant (FBP1-NLS-G260R) (N-P). The cells were then treated with 5 mM FBP for 12 h and infected with VSV (MOI, 0.1) for 10 h. Western blot analysis was used to assess FBP1 and PFKP expression (B, E, H, K and N), intracellular FBP levels were measured using a microplate reader (C, F, I, L and O), and VSV RNA was assessed by qPCR (D, G, J, M and P). siCtrl, siControl. Data are presented as the mean ± SEM. NS, not significant, * p < 0.05; ** p < 0.01; *** p < 0.001, one-way ANOVA.
Article Snippet: Primary antibodies against the following proteins were used for immunoblots: anti-HMGB1 (sc-56698, Santa Cruz Biotechnology), Anti-HCV-NS3 (ab13830, Abcam), Actin (A5541, Sigma), VSV-G(ab183497, Abcam), p-TBK1 (#5483, CST), TBK1 (#3504, CST), p-IRF3 (#4947, CST), IRF3 (#4302, CST), p-STAT1 (#10501-1-AP, ProteinTech), STAT1 (#14994S, CST), p-STAT2 (D155166-0025, Sangon Biotech), STAT2 (D261445-0025, Sangon Biotech), Tubulin (ab7792, Abcam),
Techniques: Phospho-proteomics, Transfection, Construct, Plasmid Preparation, Activity Assay, Mutagenesis, Infection, Western Blot, Expressing