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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-VHL 17 - p19 VHL p25 VHL blot: HA upplemental 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.

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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

Transcript of 80 -

Page 1: 80 -

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.

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