Fig. 1.
CAR, ASIC3, and PSD-95 form a trimolecular complex. A) COS-7 cells were double transfected with GFP-tagged PSD-95, HA-tagged ASIC3, or FLAG-tagged CAR and subjected to IP with an antibody specific for ASIC3, followed by Western blot with tag-specific antibodies. Western blot of lysates demonstrated appropriate bands indicating successful transfection. Cells were triple transfected with GFP-PSD-95, HA-ASIC3, FLAG-CAR, HA-ASICΔ4 or FLAG-CARΔ4, and subjected to IP B) with an antibody specific for ASIC3 or C) antibodies against GFP or CAR (RmcB), followed by Western blot with GFP-, AISC3-, or CAR (1605p)-specific antibodies. Western blot of B) lysates or C) mock transfected cells subjected to immunoprecipitation. Transfections are indicated at the bottom of each blot. Blots are representative of 3 experimental replicates.