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. Author manuscript; available in PMC: 2020 Dec 10.
Published in final edited form as: Cell Rep. 2019 Dec 10;29(11):3551–3563.e3. doi: 10.1016/j.celrep.2019.11.041

Figure 6. TRIM5γ Interacts with the C Terminus of HBx and Promotes HBx K95 Site Ubiquitination.

Figure 6.

(A) Diagram of mutant HBx constructs. the numbers indicate the amino acids in the HBx constructs. (B–E) 293T cells were co-transfected with the expression vectors for Flag-TRIM5γ and GST-HBx WT or mutants (B, R96E, Del; C, X1, X2, X3; D, X2a, X2b, X3a, X3b; E, X3bD, DD, X2bD) or EV as indicated. After 28 h, cells were harvested, coIP with FLAG-HBx, and cell lysates and precipitated samples were analyzed by immunoblotting with the indicated antibodies. (F) HBx WT or truncations were co-transfected with TRIM5γ or EV into HepG2 cells; 28h later, cell lysates were subjected to immunoblot analysis using the indicated antibodies. (G) Plasmids of expression of HBx WT, K90R, or K95R were co-transfected with TRIM5γ or EV into HepG2 cells; 28h later, cell lysates were subjected to immunoblot analysis using the indicated antibodies. (H) HepG2 cells were co-transfected with the expression vectors for HA-TRIM5γ, HA-Ub, and FLAG-HBx WT or mutants as indicated; 28h later, cells were harvested and coIP with FLAG-HBx, and cell lysates and precipitated samples were analyzed by immunoblotting with the indicated antibodies. Data are representative of at least three independent experiments.