Figure 7

Fyn, but not Src, affects the surface NR2B expression after D1 receptor stimulation. (A and B) Surface biotinylation of NMDA receptors in high-density PFC neurons at 14 DIV. PFC neurons at12 DIV were transfected with Fyn siRNA for 48 h. The PFC neurons at DIV 14 were treated with DMSO (0.1%, lanes 1 and 2); SKF-81297 (10 μM, lanes 3 and 4); Fyn (A) or Src (B) knockdown PFC neurons treated with DMSO (0.1%, lanes 5 and 6); or Fyn (A) or Src (B) knockdown neurons treated with SKF-81297 (10 μM, lanes 7 and 8). After surface receptor biotinylation, 20% of the lysis supernatant was used to detect the total (T) proteins. The remaining 80% of the supernatant was incubated with NeurAvidin Agarose beads and purified as surface (S) proteins. After SDS-PAGE the proteins were immunoblotted for NR2B and β-tubulin. (C and D) Quantification of surface NR2B expression. Surface NR2B was corrected by total NR2B to calculate the surface/total ratio and the control group was set to 100% for normalization. Fyn, but not Src, knockdown appeared to be effective in blocking the surface insertion of NR2B mediated by D1 stimulation (* p < 0.05, ** p < 0.01).