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Supplemental Figure 1. HA-Ub + + + Hypoxia + - + MG132 + + +. IP: IgG VHL. kDa 118 -. Ub-VHL. 80 -. 68 -. 48 -. 30 -. 25 -. IB: a-HA. blot: HA. 68 -. 48 -. 30 -. p25 VHL. 25 -. p19 VHL. 17 -. blot: VHL. - PowerPoint PPT Presentation
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80 -
68 -48 -
30 -25 -
kDa 118 -
IP: IgG VHL
HA-Ub + + +Hypoxia + - +MG132 + + +
68 -
48 -
30 -
25 -
IB: a-HA
blot: VHL
Ub-
VH
L17 - p19 VHL
p25 VHL
blot: HA
Supplemental Figure 1
786-O G7F cells were transfected with an HA-Ubiquitin expression vector. Twenty four hours later, transfected cells were treated with MG-132 10 M and with hypoxia or normoxia for 8 hours. Cells were lysed with NP40 buffer and immunoprecipitation was performed with anti-VHL (Ig32) antibody and western blot was further performed with anti-HA (Y11) (top panel) or anti-VHL (Ig32) (bottom panel), respectively.