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