FIG. 5.

Kette Tyr482 is the primary regulatory site for dAbl and PTP61F. (A) The WT and various mutant forms of Flag-tagged Kette^Myr were ectopically expressed in S2 cells. Immunoprecipitated (IP) Kette^Myr or an aliquot of total lysates was analyzed by immunoblotting (IB). (B) The WT or Y482F mutant form of Flag-tagged Kette^Myr was coexpressed with dAbl in S2 cells. Immunoprecipitated Kette^Myr or an aliquot of total lysates was analyzed by immuoblotting. (C) S2 cells were treated with dsRNA for PTP61F or green fluorescent protein (GFP). The dsRNA-treated cells were then transfected with the Y482F mutant form of Flag-tagged Kette^Myr. Immunoprecipitated Kette^Myr or an aliquot of total lysates was analyzed by immunoblotting. (D) Both WT and Y482F mutant forms of Flag-tagged Kette^Myr were coexpressed with either the WT or DA mutant form of HA-tagged PTP61Fm in S2 cells. PTP61Fm was immunoprecipitated with anti-HA antibody. The immunocomplex or an aliquot of total lysates was analyzed by immunoblotting. (E) All experimental procedures were as described for panel B with the exception of ectopic expression of Flag-tagged cytosolic Kette instead of Kette^Myr. (F) All experimental procedures were as described for panel C with the exception of ectopic expression of Flag-tagged cytosolic Kette instead of Kette^Myr.