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

Figure 5.

Figure 5.

GSK3β facilitates TRUSS degradation. Huh-7 cells were transfected with expression vectors of WT-Skp2 with increasing concentrations of GSK3β inhibitor viii (5 μM, 10 μM and 20 μM) (B). Cell lysates were western-blotted for TRUSS and GAPDH. Huh-7 cells were transiently transfected either with 2.5 μg WT-Sp2 alone or along with GSK3β inhibitor viii (20 μM) (C). After 48 h, cells were incubated with cycloheximide for indicated time points and the cell lysates were western-blotted for TRUSS, and GAPDH. Cells were transfected with wild-type HA TRUSS (1 μg), in combination with 2 μg His-Ub, WT-Skp2 (1 μg) and increasing doses of GSK3β inhibitor viii (5 μM, 10 μM and 20 μM) (D). The cell lysates were harvested after MG132 treatment and immunoprecipitated with anti-HA antibody followed by immunoblotting for His.