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. Author manuscript; available in PMC: 2014 Apr 25.
Published in final edited form as: Mol Cell. 2013 Mar 14;50(2):172–184. doi: 10.1016/j.molcel.2013.02.014

Figure 6. The Uba6-Use1 pathway promotes Ube3a turnover in MEFs.

Figure 6

(A) Uba6Flox/Flox MEFs were incubated with Adenoviral-GFP (as control) or with Adenoviral-CRE and after 48h, cell extracts were analyzed by immunoblotting using the indicated antibodies. (B) qPCR analysis of Ube3a mRNA from Control MEFs, MEFs depleted of Use1 by RNAi, and Uba6Flox/Flox f MEFS in which Uba6 was deleted with Adenoviral-CRE. Error bars are mean ± SEM, with n=3 independent experiments.

(C) Uba6Flox/Flox MEFs were incubated with Adenoviral-GFP (as control) or with Adenoviral-CRE and after 48h, cells were treated with cycloheximide (CHX). Cell extracts harvested at the indicated times were analyzed by immunoblotting using the indicated antibodies.

(D) MEFs were transfected with either control siRNA or 3 individual siRNAs targeting Use1. After 72 h, cell extracts were analyzed by immunoblotting using the indicated antibodies.