Source was cell extract of Kc167 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Chromatin was digested to primarily mono- and dinucleosomes with micrococcal nuclease prior to immunoprecipitation.

Source was cell extract of Kc167 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Chromatin was digested to primarily mono- and dinucleosomes with micrococcal nuclease prior to immunoprecipitation.

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

While the majority of cid and cal1 protein is found in chromatin fractions, a cid-cal1 interaction is detected by coimmunoprecipitation using a chromatin-free fraction. Thus cal1 binds "pre-nucleosomal" cid.

Cells were gently lysed to obtain a chromatin-free ("S1") fraction for coimmunoprecipitation.

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

cid, cal1 and His4 interact together as part of a complex.

Proteins were coexpressed in bacterial cells, affinity purified by virtue of the His tag and further purified by gel filtration.

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

cid, His4 and cal1 form a complex.

Proteins were separately purified, mixed and refolded. Complex formation was examined using gel filtration.

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

rdx binds cal1, and depletion of rdx leads to a proteasome-dependent decrease in the level of cal1 protein.

In vitro, cal1 is required for ubiquitination of cid.

Positive control.

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

Source was U2OS cells; bait produced from transfected construct; prey produced from transfected construct.

The ability of cid or cid/His3 chimera fused to LacI to recruit cal1 to LacO arrays was taken as evidence of a cid-cal1 interaction.

Source was U2OS cells; bait produced from transfected construct; prey produced from transfected construct.

The ability of cal1 fused to LacI to recruit cid to LacO arrays was taken as evidence of a cid-cal1 interaction.

Trimeric complex consisting of cal1, cid and His4 was captured as three binary interactions.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

cal1 was used as bait to pull down a cid-His4 complex.

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 S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

Bait protein was fused to lacI, which targets the chimera to transgenic chromosomal lacI binding sites. The ability of this chimera to recruit prey protein to the lacI binding sites was taken as evidence of direct physical interaction.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Trimeric complex consisting of cal1, cid and His4 was captured as three binary interactions.

Trimeric complex consisting of cal1, cid and His4 was captured as three binary interactions.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.