Journal: PLoS Pathogens
Article Title: Aerobic glycolysis supports hepatitis B virus protein synthesis through interaction between viral surface antigen and pyruvate kinase isoform M2
doi: 10.1371/journal.ppat.1008866
Figure Lengend Snippet: (A) LHBS-positive immortalized hepatocytes were treated with DMSO or 50 μΜ DASA-58 for 2 days. Total cell lysates were collected for measuring PKM2 activity. Relative PKM2 kinase activity of LHBS-positive cells was shown after normalization with that of DMSO treatment cells. (B) HuH-7 cells were transfected with pHBV3.6 for 24 hours and then treated with DMSO or 50 μΜ DASA-58 for additional 24 hours. Production of HBsAg and HBeAg in the culture supernatants were measured. Relative extracellular HBsAg and HBeAg were shown with DMSO-treated group as an internal control. (C) HuH-7 cells were transfected with pHBV3.6 for 24 hours and then treated with DMSO or 50 μΜ DASA-58 for additional 24 hours. Representative western blots of HBV viral products were shown on the left panels. The right panel showed changes of indicated viral products after DASA-58 treatment in comparison to DMSO-treated cells as the control. (D) Intracellular abundance of 6-PGA was measured in pHBV3.6-transfected HuH-7 cells following treatments of DMSO, 20 μM TEPP-46 or 50 μΜ DASA-58 for 24 hours. Quantitative results of 6-PGA relative abundance were shown. (E) The production of virions in HBV-infected HepG2-NTCP-C4 cells, with or without treatments of chemical PKM2 activators were measured by detecting HBV DNA in the culture supernatant using quantitative PCR. All quantitative results were a summary of three repeats and data were displayed as mean ± standard error. *p<0.05, **p<0.01, and ***p<0.001 were calculated using Student’s t test.
Article Snippet: TEPP-46 (ML-265) and 2-deoxy-D-Glucose (2-DG) were purchased from Cayman Chemical.
Techniques: Activity Assay, Transfection, Western Blot, Infection, Real-time Polymerase Chain Reaction
