ml 265  (MedChemExpress)

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: HuH-7 cells were transfected with pHBV3.6 for 24 hours and then treated with DMSO or 20 μΜ TEPP-46 for additional 24 hours. (A) The culture media of DMSO or TEPP-46 treatment were collected and subjected for measurement of HBsAg and HBeAg level. (B) Protein expressions of HBV viral products (left panel) and host proteins (right panel) upon treatment of DMSO or TEPP-46 were shown in western blotting. (C) Quantitative results of viral and host protein expressions were shown, with actin as an internal control. 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: Transfection, Western Blot

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

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) Colony formation assay was performed with control and LHBS-positive immortalized hepatocytes for 3 weeks in soft agar plates. Representative plate images are shown. No colony formation was detected in control cells. Quantitation of colony numbers were provided in the right panel. (B) LHBS-positive hepatocytes were treated with DMSO or 20 μΜ TEPP-46 for 24 hours and cell lysates were harvested for measuring PKM2 activity. (C) Cell viability was measured in LHBS-positive hepatocytes 48 hours after treatments with DMSO or 20 μΜ TEPP-46. (D) Representative colony formation images of control, and LHBS-positive hepatocytes treated with DMSO or 20 μΜ TEPP-46 were shown. Quantitation results of colony numbers were shown on the right panel. (E) Representative colony formation images of control, and LHBS-positive hepatocytes treated with DMSO or 50 μΜ DASA-58 were shown. Quantitation results of colony numbers were shown on the right panel. All quantitative results were a summary of three repeats and data were displayed as mean ± standard error. *p<0.05 was 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: Colony Assay, Quantitation Assay, Activity Assay