Crk-I and Crk-II coimmunoprecipitate with GCKR. HEK 293T cells were transfected with constructs that directed the expression of HA-GCKR and Crk-I or Crk-II.

Lysates prepared from the transfected cells were immunoprecipitated with HA, FLAG (control antibody), or a Crk antibody as indicated. The immunoprecipitates were fractionated on SDS-PAGE, transferred, and blotted with the HA antibody (top) or the Crk antibody (bottom). Locations of HA-GCKR, Crk-II, and Crk-I are indicated with arrows as the immunoglobulin light chains. These experiments were performed 3 times with similar results.