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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: J Neurochem. 2011 Mar 17;119(1):27–39. doi: 10.1111/j.1471-4159.2011.07221.x

Figure 7.

Figure 7

Nedd4 ubiquitinates AMPARs and suppresses receptor surface expression in neurons. (A) HEK cells were first transfected with GluA1 and HA-ubiquitin cDNA plasmids and then incubated with lentiviral Nedd4 and a viral vector control. GluA1 immunoprecipitates were probed with anti-ubiquitin antibodies (left panel). Nedd4, GluA1 and tubulin were probed in cell lysates (right panel). (B) Neurons were infected with viral Nedd4 and ubiquitination of endogenous GluA1 was examined (left panel). Pooled data are shown in the bar graph (right panel, n=3). (C) Neurons were transfected with Nedd4 siRNA or scrambled control siRNA, with a subsequent incubation with the viral ubiquitin construct. Nedd4 siRNA suppressed Nedd4 expression and reduced GluA1 ubiquitination (n=3). (D and E) Neurons were transfected with GFP plus Nedd4, or GFP alone as a control. Surface (Surf) and total GluA1 were detected sequentially (n=19 cells in control and n=17 in Nedd4, from two independent assays). Bar graph data represent means ± SEM, * P<0.05, t test. Scale bar, 20 μm.