To evaluate the surface expression of GluA1 using immunofluorescence microscopy we cultured dissociated rat hippocampal neurons transfected with CNIH-2 shRNA and visualized somatic and dendritic surface GluA1 immunoreactivity ~20 days later. CNIH-2 shRNA transfected neurons were compared to adjacent untransfected neurons. CNIH-2 knock-down dramatically reduced surface GluA1 (Figures 4A and S5A), consistent with our findings showing reduction of synaptic currents. Transfection of a scrambled shRNA or GFP alone had no effect on surface GluA1 staining (Figures 4B and S5B–C).
