Figure 4.
Leptin-regulated NR2BY1472 phosphorylation and surface expression is Fyn dependent. (A) Representative Western blot of HEK293T cells transfected with NR2B-V5, NR1, LepRb-myc, and either V5-Fyn or V5-DN Fyn and treated with leptin (50 nM, 2 hours). (B) Quantification of pNR2BY1472 intensity normalized to total NR2B-V5 intensity (n = 3). (C) Hippocampal neurons were transfected with Clover and EGFP-NR2B-V5 and either V5-Fyn or V5-DN Fyn ± leptin stimulation (50 nM, 2 hours) and live immunostained for surface EGFP–NR2B. Quantification of immunostained EGFP-integrated signal density (n = 15). All experiments were repeated in 3 independent culture preparations and expressed as the mean ± SEM, *P < 0.05, **P < 0.01.
Abbreviations: DN, dominant negative; pNR2B, phospho NR2B; Y1472, tyrosine 1472.