Protein expression of GluN3A in whole brain, 46C ESCs, non-transfected NSCs, and GluN1-transfected NSCs.
A: GluN3A expression in total protein preparations. Bands at the expected molecular weight of GluN3A (130 kDa) were visible in the positive control (mouse whole brain P3) and in non-transfected 46C-derived NSCs. Undifferentiated 46C ESCs did not express GluN3A proteins. B: Expression of the housekeeping protein calnexin (90 kDa) as a control. C: GluN3A expression in total and plasma membrane protein preparations from GluN1-transfected 46C-derived NSCs. GluN3A protein expression was not detectable in the plasma membrane fraction of 46C-derived NSCs. GluN1 (130 kDA) surface expression in GluN1-transfected 46C-derived NSCs in the same total and plasma membrane protein preparations is shown to demonstrate that the biotinylated concanavalin A-treatment of NSCs successfully enriched plasma membrane protein. D: 46C-derived NSCs were immuncytochemically stained for GluN3A (red fluorescence), and cell nuclei were stained with Hoechst 33342 (blue fluorescence). The staining confirmed the expression of GluN3A in the cytoplasm of 46C-derived NSCs. Scale bars: 50 μm.