Effects of high-fat diet treatment on Fyn-mediated NR2B activation (n = 10/group). To explore the putative mechanisms underlying the effects of high-fat diet treatment on prediabetic neuropathy, spinal cord tissues were collected from each mouse at 16 wk or 24 wk in wild-type mice. Western blot analysis was performed on total NR2B levels (A), pNR2B-to-NR2B ratio (B), and Fyn kinase levels (C). D: to study physical interaction between Fyn kinase and NR2B, coimmunoprecipitation (Co-IP) assays were conducted by immunoprecipitation of the spinal cord samples with mouse monoclonal anti-Fyn antibody, followed by probing the blots with the antibody against NR2B (WB). *Significant difference relative to normal diet control (P < 0.05); #significant difference relative to 16 wk (P < 0.05).