Figure 3.

Reverse immunoprecipitation of suspected targets. Antibodies against putative interacting proteins were used for immunoprecipitations from rat cortical lysates (500 μg protein). Proteins were extracted from the Protein-A agarose beads without boiling to prevent formation of large molecular weight aggregates of GLT-1 (Haugeto et al., 1996). The bottom row of immunoblots shows the putative target, and the top row shows GLT-1 immunoreactivity (marked with arrowheads). In some cases, the putative interacting protein could not be resolved from IgG dimers (∼100 kDa). In these cases (marked with asterisks), parallel immunoprecipitations were conducted, and the putative interacting protein was extracted with boiling in sample buffer so the IgG migrates at 50 kDa. These data are representative of at least three independent experiments.