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).

Various stretches of six to seven arm repeats can bind to shg, but smaller fragments do not.

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

CadN binds the smaller 82 kDa arm isoform, whereas shg prerentially binds the 105 kDa arm isoform.

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

Glycoproteins coimmunoprecipitating with arm were detected by concanavalin A staining of the blot.

Positive control.

Bait endogenous to embryonic extract.

Prey endogenous to embryonic extract.

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

Positive control.

The interaction of arm protein with the cytoplasmic domain of the shg protein is not affected by mutations to the JM region that abolish the shg-p120ctn interaction.

Source was embryos of transgenic fly line; bait produced from endogenous gene in transgenic embryos for a dominant negative form of Src42A[Kr] or a constitutively active form Src42A[YF].; prey produced from endogenous gene.

interaction seen only in membrane fractions. No difference between wild-type and transgenic embryo extract

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

Binding of arm to shg in extracts derived from early embryos (not late embryos) was disrupted by mildly acidic conditions (pH5.5). This suggests that binding of arm to shg is weaker in early embryos than in later embryos.

Immunoprecipitated complexes from early and late embryo extracts were washed with mild acidic solution (pH5.5).

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

Interaction stronger in a Btk29A mutant background.

Positive control.

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

Source was embryos of transgenic fly line; fluorescence donor produced from tagged transgenic construct; fluorescence acceptor produced by injected antibody recognition of endogenous gene product.

Changes in the strength of the arm-shg interaction during mesoderm invagination or experimentally induced mechanical strain inferred from changes in donor fluorescence lifetime. Close proximity of the fluorescence donor and acceptor leads to diminished donor lifetime, thus an increased donor lifetime suggests an increase in distance between arm and shg during mesoderm invagination.