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. Author manuscript; available in PMC: 2013 Nov 30.
Published in final edited form as: Cell Physiol Biochem. 2012 Nov 30;30(6):1444–1455. doi: 10.1159/000343332

Fig. 2.

Fig. 2

Interaction between NBCn1 and PSD-95. A) Pull-down assay. Rat brain lysates were incubated with either the GST/NBCn1 fusion protein containing the C-terminal 131 amino acids of NBCn1, GST/NBCn2 containing the C-terminal 113 amino acids of NBCn2 (NCBE), or GST/NBCn1-ETSL that lacks the last 4 amino acids of NBCn1. GST only served as a control. Pull-down samples were immunoblotted with the PSD-95 antibody. A weak band of ~50 kDa is nonspecific. Coomassie blue staining of the protein gel showed GST fusion proteins with the expected molecular weight. B) Coimmunoprecipitation. Lysates of HEK 293 cells expressing NBCn1/PSD-95 or NBCn1 alone were immunoprecipitated with the PSD-95 antibody and then immunoblotted with the NBCn1 antibody. C) Coimmunoprecipitation of NBCn1/PSD-95 and PSD-95/NR2A. Lysates of cells expressing NBCn1, NR1A/NR2A (i.e., NMDAR), and either PSD-95 or none were immunoprecipitated with the PSD-95 antibody and then immunoblotted with antibodies to NBCn1 and NR2A. The amount of the total protein in the lysates corresponds to one fifteenth of that in the precipitates. L, lysates; P, immunoprecipitates. D) Competition analysis of NBCn1/PSD-95 and PSD-95/NR2A. Cells were transfected with PSD-95, NMDAR and different amounts of NBCn1 (0.5–4 μg).