Interaction in vitro; bait derived from transfected COS cells; prey purified from medium of transfected S2 cells.

robo-transfected COS cells wre incubated with supernatants from sli-transfected S2 cells. Binding to the cell surface was determined by immunofluorescence using sli antibody.

Interaction in vitro; bait derived from transfected COS cells; prey purified from medium of transfected COS cells.

sli and robo bind with a K[[d]] = 41.7 nM.

sli-transfected COS cells wre incubated with purified Fc-tagged robo, then with [125]I anti-human IgG. Total binding to the cell surface was determined by measuring cell-associated radioactivity.

Interaction in vitro; bait purified from conditioned medium of transfected 293-EBNA cells; prey purified from conditioned medium of transfected 293-EBNA cells.

The robo1-sli interaction is enhanced by heparin.

Interaction in vitro; bait purified from conditioned medium of transfected 293-EBNA cells; prey purified from conditioned medium of transfected 293-EBNA cells.

The robo1-sli constructs form a binary complex with 1:1 stoichiometry. A ternary 1:1:1 complex is formed with a purified heparin-derived decasaccaharide.

Interaction in vitro; bait purified from conditioned medium of transfected 293 cells; prey purified from conditioned medium of transfected 293 cells.

Mutations in robo1 Ig domain 2 have only a marginal effect on sli binding.

Interaction in vitro; bait produced as a recombinant fusion protein in baculovirus system; prey produced as a recombinant fusion protein in baculovirus system.

sli binds robo (Kd = 235nM) more strongly than lea/robo2 (Kd = 1098nM).

Positive control.

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

Interaction in vitro; bait present on live, transfected S2R+ cells; prey produced from conditioned media of transfected S2R+ cell line.

S2R+ cells expressing robo1 were incubated with sli-conditioned medium, then fixed and analyzed for sli-surface binding by immunofluorescence.

Interaction in vitro; bait present on live, transfected S2R+ cells; prey produced from conditioned media of transfected S2R+ cell line.

Only robo1 Ig1 domain is necessary for sli binding (other Ig domains tested are dispensable).

S2R+ cells expressing robo1 were incubated with sli-conditioned medium, then fixed and analyzed for sli-surface binding by immunofluorescence.

Source was cell extract of 293 cell line; bait produced from transfected construct; prey produced from transfected construct.

In the absence of Dscam1, robo1 preferentially interacts with full length sli. Upon cotransfection of Dscam1, robo1 preferentially binds the proteolytically processed sliN form.

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

The sli-robo1 interaction occurs in both wildtype and mmy mutant backgrounds.

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

Interaction is detected in wildtype and in ptc mutant embryos.

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

Precipitation of robo1 coprecipitates full length sli,the processed N-terminal fragment of sli but not the C-terminal fragment.

The interaction was observed on the surface of SR2+ cells expressing robo1. sli was derived from the medium of transfected S2R+ and bound sli was detected with anti-sli antibodies.

Source was S2R+ cell line; bait produced from transfected construct; prey produced from transfected construct.