Figure 1.

Recombinant and endogenous gephyrin contains consensus sequences for proline-directed phosphorylation. (A) HEK 293 cells transfected with gephyrin-FLAG were lysed, half of the lysate was treated with CIP and immunoprecipitated with anti-FLAG monoclonal antibody (lanes 4 and 6) and with a non-related antibody (anti-MYC) as negative control (lane 2). Immunoprecipitates were analysed by Western blotting using the MPM-2 antibody (left panel). The same membrane was stripped and reprobed with the anti-gephyrin antibody (right panel). (B) Endogenous gephyrin was immunoprecipitated from mouse brain lysate with an anti-gephyrin polyclonal antibody (lanes 4 and 6) and with normal rabbit serum (NRS) as negative control (lane 2). Half of the cell homogenate was treated with CIP before immunoprecipitation. Immunoprecipitates were analysed as described in (A).