Two-hybrid system: yeast

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from embryo cDNA expression library).

raps coimmunoprecipitates with Gαi in the presense of GDP, but not in the presence on non-hydrolyzable GTPγS.

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

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 embryos of transgenic fly line; prey produced from bait produced from endogenous gene.; bait produced from endogenous gene.

Transgenic embryos conaining a heat shock inducible allele of loco were used in this experiment. Heat shock treatment did not have an effect on pins binding to Gαi.

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

pins does not interact with Gαi if Gαi is bound to GTP.

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

Similar experiments done in a ric8a mutant background demonstrated the requirement of ric8a for interaction between pins and Gαi.

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

In this experiment, pins dos not interact with GTP bound Gαi.

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

Gαi precipitates endogenous pins from S2 cells in this experiment.

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

pins binds Gαi only in the presence of GDP, not GTP.

positive control

Bait expressed in transiently transfected cells.

Prey expressed in transiently transfected cells.

positive control

This interaction occurs in wildtype and Tre1 mutant embryos.