Upon expression of hh protein, phosphorylated forms of fu protein preferentially coimmunoprecipitate with cos protein.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

fu and cos proteins cofractionate in a complex with an apparent molecular weight of 700 kDa.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

fu and cos proteins coimmunoprecipitate from high molecular weight complexes fractionated by gel filtration.

Source was embryos of wild-type fly line; bait produced from endogenous gene; prey produced from endogenous gene.

Source was imaginal discs of wild-type or mutant fly line; bait produced from endogenous gene; prey produced from endogenous gene.

fu-cos interaction also occurs in a Su(fu) mutant background.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.

cos does not bind to Su(fu) in this experiment.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/B42-AD, Neither full-length cos nor full-length fu, nor the catalytic domain of fu (aa 1-305) could be used in this assay due to interfering activities.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was larval wing discs of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

Bait protein was fused to the motor domain of kinesin heavy chain, which targets the chimera to the plus end of microtubules. The ability of this chimera to recruit prey protein to the plus end of microtubules was taken as evidence of direct physical interaction.

Positive control.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

Bait expressed in transiently transfected cells.

Prey expressed in transiently transfected cells.

Source was cell extract of CME-W1-Cl.8+ cell line; bait produced from endogenous gene; prey produced from endogenous gene.

Cells treated with hh.

Source was cell extract of CME-W1-Cl.8+ cell line; bait produced from endogenous gene; prey produced from endogenous gene.

Cells treated with hh.

This interaction was observed in the presence or absence of hh expression.

Source was cell extract of CME-W1-Cl.8+ cell line; bait produced from endogenous gene; prey produced from endogenous gene.

The interaction between fu and cos is increased in the presence of hh, smo and Gprk2.

Source was live S2 cells; proteins produced from transfected constructs.

Full-length fu was co-expressed with C-terminal fu and cos. With increasing concentrations of cos, the dimerization of C-terminal and full-length fu was abolished.

Interaction in vitro; bait and prey produced as recombinant fusion proteins in baculovirus system.

Interaction in vitro; bait and prey produced as recombinant fusion proteins in baculovirus system.

The kinase domain of fu only interacts with cos with the addition of the C-terminal domain of fu.