Adult mouse hippocampi were homogenized and the nuclear pellet was removed by centrifugation and resuspended in 1% Triton X-100. Pre-cleared lysates were incubated with antibody-bound Sepharose beads (Sigma). Beads were washed with lysis buffer and analyzed by immunoblotting with the relevant antibodies as indicated. For glycosylation analysis, the precleared lysate was immunoprecipitated with GluA1 or GluA2 antibody, and treated with endoglycosidase Hf (Endo H) or PNGase F overnight at 37°C, resolved by SDS-PAGE and analyzed by immunoblotting with indicated antibodies.
