Figure 5
D1 agonist induces no changes in NR2B expression after Fyn knockdown. Immunostaining revealed that neither NR2B nor PSD95 expression was changed by treatment with SKF-81297 (10 μM) in Fyn siRNA-transfected PFC neurons (d, e, f) compared with control (a, b, c). Scale bar = 10 μm. (B) Quantification of total NR2B immunofluorescence staining revealed that SKF-81297 (10 μM) had no clear effects on NR2B puncta number or fluorescence intensity in the dendrites of Fyn siRNA-transfected neurons compared with control group (p > 0.05). (C) Total protein levels of NR2B, which were determined by Western blotting after Fyn siRNA transfection followed by SKF treatment in cultured PFC neurons, were consistently unaltered by treatment with SKF-81297 (10 μM, p > 0.05) although SKF-81297 (10 μM) induced a significant increase in NR2B expression in the control condition (*p < 0.05). Similarly, in neurons with Fyn siRNA, NR2B Tyr1472 phosphorylation level was not changed by treatment with SKF-81297 (10 μM, p > 0.05). Integrated intensity analysis was performed using Image J software and the control protein level of NR2B or the Tyr1472 phosphorylation was set to 100% after being normalized by loading control tubulin.