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. 2016 Jun 20;291(34):17557–17568. doi: 10.1074/jbc.M116.731745

FIGURE 5.

FIGURE 5.

Simultaneous deletion of the middle loop and C terminus is required to block binding of Smurf2 and Itch. Full-length mPtch1 as well as the ΔML, ΔC, and ΔMLΔC mutants were transiently transfected into HEK 293 cells with FLAG-tagged Itch (A) or FLAG-tagged Smurf2 (B). α-FLAG immunoprecipitations (IP) demonstrate that the ΔML and ΔC mutants are capable of binding to both Itch and Smurf2, whereas the ΔMLΔC mutant is unable to bind to these WW domain-containing ubiquitin ligases. IB, immunoblot.