Src and Fyn knockdowns have no effect on the overall expression or localization and clustering of NR2B subunit at basal conditions. (A) 11 DIV PFC neurons were transfected with Fyn siRNA for 48 h and stained for endogenous NR2B (green) and PSD95 (red). Scale bars = 10 μm. (B) 11 DIV PFC neurons were transfected with Src siRNA for 48 h and stained for endogenous NR2B (green) and PSD95 (red). Scale bars = 10 μm. (C) Quantification of total NR2B immunofluorescence staining showed that neither Fyn nor Src had a detectable effect on total NR2B puncta number or immunofluorescence intensity compared with the control group (p > 0.05). Data represent mean ± SEM. (D) Total protein levels of NR2B were determined by Western blotting after Fyn and Src siRNA transfection in cultured PFC neurons. The blots were stripped and reprobed with antitubulin as loading control. Control protein levels of NR2B were set at 100% after being normalized to loading control tubulin (p > 0.05).
Hu et al. Molecular Brain 2010 3:20 doi:10.1186/1756-6606-3-20