FIGURE 4.

CD300c and CD300b interact through their Ig-like domains. COS-7 cells were transiently transfected with HA-tagged CD300c in combination with FLAG-tagged CD300b. Wild type forms were tested against transmembrane substitution mutants (A), intracellular deletion mutants (B), extracellular deletion mutants (C), cysteine substitution mutants (D), or point mutants affecting the protruding body in the Ig fold (E) to map the interaction between both molecules. Cells were lysed (1% Triton X-100) and CD300c was immunoprecipitated (IP) with anti-HA.11 mAb in all cases. Western blots (WB) were conducted with the indicated Abs. Whole cell lysates (2%) were included as controls.