Immunoprecipitation of phosphorylated GluA1 from denatured hippocampal homogenates. (A) Example immunoblots showing total and phosphorylated GluA1 in immunoprecipitated (left) and unbound fractions (right) following immunoprecipitation of T840-phosphorylated GluA1. (B) Plots show levels of total and phosphorylated GluA1 in the unbound fraction following immunoprecipitations using either phospho-T840 GluA1 or control IgG antibodies (n = 5, lines connect results from the same homogenate). GluA1 levels in the unbound fraction were reduced to 51 ± 9% of input following immunoprecipitation of T840-phosphorylated GluA1 (P < 0.01 compared with levels in IgG control unbound fraction) (C,D) Same as in panels A and B except immunoprecipitations were performed using phospho-S845 GluA1 antibodies (n = 4). GluA1 levels in the unbound fraction were 98 ± 8% of the input following immunoprecipitation of S845-phosphorylated GluA1 (P = 0.41 compared with levels in IgG control unbound fraction). Bars in point plots indicate median values.
