Coimmunoprecipitation of CrkL with GCKR depends on the region in GCKR between amino acids 599 and 699.

HEK 293T cells were transfected with constructs that directed the expression of CrkL and GCKR or truncated versions of GCKR. Cell lysates and myc (control antibody) and HA antibody immunoprecipitates were analyzed by immunoblotting. The blot was sequentially reacted with the HA antibody (top) and the CrkL antibody (bottom). CrkL is indicated by an arrow. The truncated GCKR proteins can be seen in the cell lysate and in the HA immunoprecipitation lanes (arrows). Several nonspecific proteins are also present (top), the most prominent just below 95 kd in the cell lysate lanes.