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

Sas-4-containing complexes were affinity-purified, then subjected to immunoprecipitation by antibodies to asl protein.

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

Coimmunoprecipitation from "high-speed lysates" depleted of mature centrosomes.

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

Coimmunoprecipitation from "high-speed lysates" depleted of mature centrosomes.

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

Characterization of Sas-4 complexes shows that Sas-4 and asl cosediment in a narrow range of fractions at 7S.

Coimmunoprecipitation from "high-speed lysates" depleted of mature centrosomes.

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

asl coimmunoprecipitates and co-fractionates with Sas-4 as part of the S-CAP complex (centrosomal scaffold).

Sas-4 complexes were immunoprecipitated from "high-speed lysates" depleted of mature centrosomes and different Sas-4-containing complexes were fractionated on a sucrose gradient.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey derived from wild-type embryonic extract.

Free tubulin can disrupt the interaction of Sas-4 with asl; mutation of the Sas-4 tubulin binding domain prevents this competition.

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

The interaction of Sas-4 with asl is disrupted by the addition of a non-hydrolyzable GTP analog, GMPCPP, to the extract.

The interaction of Sas-4 with asl is increased by the addition of griseofulvin to the extract, a compound that induces hydrolysis of tubulin-bound GTP to GDP.

Coimmunoprecipitation from "high-speed lysates" depleted of mature centrosomes.

This interaction is not affected by mutations to the β9-10 surface of the TCP domain: E792A, V800A/I802A or L841K.

Source was cell extract of C131 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Constructs were expressed in acentrosomal cell line derived from Sas-4 null homozygous mutant flies.

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

Two-hybrid system: yeast GAL4-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 yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).