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. 2015 Jun 3;14(16):2688–2700. doi: 10.1080/15384101.2015.1056946

Figure 6.

Figure 6.

Stabilization of TRUSS in the presence of viral HBx. Huh-7 cells were co-transfected with the HBx expression vector X0 and HA-TRUSS (1.5 µg each) (A) or X0 alone (B) and the cell lysates were protein gel blotted for TRUSS, c-Myc and HBx. (C) Cell lysates of un-transfected HepG2 and HepG2.2.15 cells were western-blotted for endogenous TRUSS. Huh-7 cells were transfected with X0 (2.5 µg) along with 2.5 µg each of either wild-type HA-TRUSS (D) or TRUSS deletion mutants (E), and the cell lysates were immunoprecipitated with anti HA antibody followed by immuno-blotting for HBx or recombinant TRUSS. Huh-7 cells were transfected with expression vectors HA-TRUSS (1 µg) along with the expression vectors of HBx (2 µg) (set I) or 2 µg each of HBx and Skp2 (Set II) (F), the cell lysates were immunoprecipitated with anti HA antibody followed by immuno-blotting for HBx. (G) For ubiquitination assay, cells were transfected with HA-TRUSS in combination with Ub and or X0 and the cell lysates were harvested after MG132 treatment and immunoprecipitated with anti-HA antibody for the recombinant TRUSS followed by immuno-blotting with anti-His antibody.