Fig. 1.

UCP degraded missense mutant pVHLs (V155A, L158Q and Q164R) via proteasome pathway and decrease half-life in cell. a HEK293T cells were transfected with HA-tagged mutant VHL (V155A, L158Q, Q164R) and/or Flag-UCP and cells were incubated for 16 h either in the presence or absence of 10 uM MG132 at 36 h post- transfection. b Mutants VHL were transfected into 293 T cell. At 36 h post- transfection. Cells were treated with cyclohexamide (CHX) for 0, 2, 4, 6 h then and immunoblotted as indicated. c 786-O cell lines constitutively expressing mutant pVHL were treated with cyclohexamide (CHX) for 0, 2, 4, 6 h then and immunoblotted as indicated. Calculation of protein degradation kinetic of both HEK293T (d) and 786-O (e) cell lines was revealed that mutant pVHLs have shortened half-life (n = 3, p < 0.01). f HEK293T cells were transfected with HA-tagged mutant VHL and/or UCP-shRNA and then at 48 h post- transfection, cell were harvested and immunoblotted as indicated