Source was cell extract of S2 cell line; bait and prey produced from transfected constructs.

An enzymatically inactive Smurf construct and an un-tagged, constitutively active construct of tkv was used. Two-step coIP demonstrating Smurf precipitates IntS8 and tkv in the first step. The first IP reactions was used for a second IntS8 coIP which demonstrated an interaction with Smurf and tkv, suggesting all three proteins associate.