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

7 sd clones were recovered in the yki screen.

Two-hybrid system: yeast Stratagene CytoTrap

Interaction of prey with bait targets hSos domain to membrane, activating Ras and permitting mutant cdc25H yeast to grow at 37oC.

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

vg outcompetes yki for binding to sd when the three proteins are coexpressed.

The majority of yki protein is cytoplasmic. To avoid isolating these cytoplasmic factors and instead identify proteins involved in yki transcriptional functions, yki was immunoprecipitated from nuclear extracts.

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

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

The sd-yki interaction is not affected by the presence of wild-type Tgi. However, a Tgi variant carrying three disabled PPXY motifs does disrupt the sd-yki interaction.

FRET efficiency between sd and yki was decreased upon cotransfection of Tgi.

Source was live S2 cells; bait produced from transfected construct; prey produced from transfected construct.

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

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

Positive control.

Sequential coimmunoprecipitation, first of FLAG-Tgi, then of Myc-yki, in the presence of HA-sd, suggests that sd, yki and Tgi form a ternary complex.

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

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

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

Positive control.

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

Mutant form of yki S168A used to show

Increased expression of Tgi results in a decrease in binding between sd and yki in a dose-dependent manner.

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

Increased expression of vg results in a decrease in binding between sd and yki in a dose-dependent manner.

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

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

The sd-yki interaction decreases upon coexpression of E2f1.

Source was cell extract of HEK293T cell line; bait produced from transfected construct; prey produced from transfected construct.

The sd-yki interaction decreases upon coexpression of E2f1.

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

Coexpression of CG7028 reduces the yki-sd interaction.

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

Co-expression of the vg TONDU domain peptide inhibits the yki-sd interaction.

No difference in yki-sd interaction with different yki isoforms including or excluding exon 3.

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

9.5 +/- 0.5 nM

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

Source was cell extract of HEK293T cell line; bait produced from transfected construct; prey produced from transfected construct.

foxo but not foxo-Mu (322PPPY-AAAA325) transfection diminished yki-sd interaction.

Source was cell extract of HEK293T cell line; bait produced from transfected construct; prey produced from transfected construct.

foxo but not foxo-Mu (322PPPY-AAAA325) transfection diminished yki-sd interaction. Addition of hydrogen peroxide or Paraquat diminished yki-sd interaction.