Fig. 5
RIM1 is a binding partner of the NMDAR complex in mouse brain. a RIM1 is within the NMDAR complex, but not the AMPAR complex. Extracts from adult mouse cortex were immunoprecipitated with antibodies against GluN1 (left) or RIM1 (middle: antibody from ABNOVA; right, antibody from SYSY) and blotted with corresponding antibodies. IP immunoprecipitation. b RIM1 has no interaction with NMDAR subunits in transfected HEK293T cells. Extracts of HEK293T cells transfected with HA-RIM1 and GFP-GluN1 (left), GFP-GluN2A (upper right), or GFP-GluN2B (lower right) were immunoprecipitated with GluN1 or HA antibody and blotted with the corresponding antibodies. c Extracts of HEK293T cells co-transfected with HA-RIM1, GFP-GluN1, and GFP-GluN2A (upper), or HA-RIM1, GFP-GluN1, and GFP-GluN2B (lower) were immunoprecipitated with GluN2A or GluN2B antibody and blotted with the corresponding antibodies. d RIM1 interacts with PSD-95 in transfected HEK293T cells. e GluN2B interacts with PSD-95 in transfected HEK293T cells. f Extracts of HEK293T cells co-transfected with HA-RIM1, GFP-GluN2B, and mCherry-PSD-95 were immunoprecipitated with GluN2B antibody and blotted with the corresponding antibodies. g Different RIM1 fragments tagged with GFP (GFP-RIM11-705 or GFP-RIM1706-1615) were cotransfected with mCherry-PSD-95 into HEK293 cells and the interaction between these two proteins was tested by immunoprecipitation
