Figure 1
From: Direct interaction between GluR2 and GAPDH regulates AMPAR-mediated excitotoxicity
Identification and characterization of GluR2/GAPDH interaction. A, Coomassie blue stained SDS-PAGE gel of the protein(s) selectively affinity pulled down by GST-GluR2NT, GluR1NT and GST alone from solubilized rat hippocampal lysates. Protein of interest: ~37 kDa. B, Western blot analysis of rat hippocampal proteins affinity purified by GST-GluR2NT, GST-GluR2CT and GST from solubilized rat hippocampal lysates and immunoblotted with primary antibody against GAPDH. C, Co-immunoprecipitation of GAPDH by the GluR2 primary antibody from solubilized rat hippocampus. D, Schematic representation of GST-fusion proteins encoding truncated GluR2NT segments. E-G, Western blot analysis of rat hippocampal proteins affinity purified by (E) GST-GluR2NT1, GST-GluR2NT2 GST-GluR2NT3 and GST; (F) GST-GluR2NT1-1, GST-GluR2NT1-2, GST-GluR2NT1-3, GST-GluR2NT1-4, GST-GluR2NT1-5 and GST; (G) GST-GluR2NT1-3–1, GST-GluR2NT1-3–2 and GST from solubilized rat hippocampal lysates and immunoblotted with primary antibody against GAPDH. H-J: Using an in vitro binding assay, [35 S]-GAPDH probe bound with GST-GluR2NT1(H), GST-GluR2NT1-3(I) and GST-GluR2NT1-3–2(J), but not with other GST fusion proteins or GST alone.