Journal: Journal of Virology

Article Title: Ubiquitin-dependent proteasomal degradation of small hepatitis B virus surface antigen mediated by TRIM21 and antagonized by OTUD4

doi: 10.1128/jvi.02309-24

Figure Lengend Snippet: OTUD4 and TRIM21 alter the production of subviral particles and virions. (A, B, E, F) Lysates were prepared from Huh7 and HepG2 cells co-transfected with plasmids encoding 1.2HBV and either TRIM21-myc ( A, B ) or OTUD4-myc ( E, F ). These lysates were subjected to immunoblotting using antibodies against myc, β-actin, and SHBs, and intracellular HBV DNA levels were quantified by qPCR. Additionally, cell culture supernatants were analyzed by qPCR to quantify extracellular HBV DNA and virion DNA levels. (C, D, G, H) Lysates from cells transfected with plasmids encoding 1.2HBV and siRNA targeting either TRIM21 ( C, D ) or OTUD4 ( G, H ) were similarly processed. The lysates were immunoblotted using the indicated antibodies, and intracellular HBV DNA was quantified by qPCR. qPCR was performed on the supernatants to measure HBV DNA and virion DNA levels.

Article Snippet: The human hepatoma cell lines HepG2 and Huh7 were sourced from the American Type Culture Collection (Manassas, VA, USA) and the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), respectively, while HUVECs were generously provided by Dr. Huang ( ).

Techniques: Transfection, Western Blot, Cell Culture

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (B and C) HepG2.2.15 cells and HepG2 cells at the same cellular concentration (1 × 10 6 cells) were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively. (E and F) After the gradient transfection of pBlue-HBV 1.3-fold plasmids into HepG2 cells, the cells were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively, with the mRNA expression level of GAPDH as a reference. (H and I) Huh7 cells were analyzed in the same way as HepG2 cells. (A, D, and G) ELISA results for HBeAg and HBsAg concentrations in cell supernatants demonstrated successful infection.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Concentration Assay, Cell Culture, Expressing, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Infection

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (A and D) HBV 1.3-fold plasmids and various concentrations of PcDNA3.1–3*Flag-QPCT were cotransfected into Huh7 and HepG2 cells and cultured for 48 h, supernatants and cells were harvested. (A and D) The overexpression of Flag-QPCT in the transfected HepG2 and Huh7 cells was detected by Western blot. (B and E) The expression levels of HBeAg and HBsAg in the supernatants were detected by ELISA. (C and F) Total RNA was extracted from the cells, and the mRNA expression of HBV pgRNA in the cells was detected by RT‒PCR, with the mRNA expression level of GAPDH as a reference. (G and H) Total DNA was extracted from the cells, and the HBV-DNA copies in Huh7 and HepG2 cells were detected by RT‒PCR.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Cell Culture, Over Expression, Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay