Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2016 Apr 23;7(21):31440–31453. doi: 10.18632/oncotarget.8951

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright: © 2016 Qiu et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PMC Copyright notice

A-B. NEDL2 positively regulates PI3K-Akt pathway in a ubiquitin ligase activity-independent manner. HEK293T cells were transfected with or without HA-Ub and NEDL2/NEDL2(C1540A) mutant plasmids. Ten hours before harvest, the cell were treated with MG132, and then cell lysates were prepared and immunoprecipitated with anti-Flag antibody. The immunoprecipitates were analyzed by immunoblotting with the indicated antibodies (A). MCF7 cells were transfected with or without NEDL2/NEDL2(C1540A) mutant plasmids. Cells were serum-starved overnight and either untreated or treated with GDNF (40 ng/mL, 15min); then analyzed by immunoblotting with the indicated antibodies (B). (C-F) NEDL2 positively regulates PI3K-Akt pathway in a Nedd8 ligase activity-dependent manner. C. HEK293T cells were transfected with Myc-Nedd8/Myc-Nedd8 I44A mutant/Myc-Nedd8ΔGG mutant and NEDL2 mutant plasmids, and then cell lysates were prepared and immunoprecipitated with anti-Flag antibody. The immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. D. HEK293T cells were transfected with Myc-Nedd8 and the indicated NEDL2 WT and mutant plasmids, and then cell lysates were prepared and immunoprecipitated with anti-Flag antibody. The immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. E. HEK293T cells were transfected with HA-Ub and NEDL2 plasmids, and then cell lysates were prepared and immunoprecipitated with anti-Flag antibody. The immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. F. MCF7 cells were transfected with or without NEDL2/NEDL2(C1341A) mutant plasmids. Cells were serum-starved overnight and either untreated or treated with GDNF (40 ng/mL, 15 min) then analyzed by immunoblotting with the indicated antibodies.