Bait endogenous to embryonic extract.

Prey derived from a transgenic line carrying a HA-tagged esc construct, P{HA-esc}.

Bait derived from a transgenic line carrying a HA-tagged esc construct, P{HA-esc}.

Prey endogenous to embryonic extract.

Bait expressed and purified from bacteria.

Prey derived from in vitro translation.

GST pull down.

Bait expressed and purified from bacteria.

Prey derived from in vitro translation.

GST pull down.

Bait expressed and purified from bacteria.

Prey derived from a transgenic line carrying a HA-tagged esc construct, P{HA-esc}.

GST pull down.

Bait expressed and purified from bacteria.

Prey endogenous to embryonic extract.

GST pull down.

Interaction in vitro; bait produced and labeled by in vitro translation; prey produced and labeled by in vitro translation.

The entire WD repeats region of esc is required for binding to E(z). Many small deletions throughout this region disrupt binding (data not shown).

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.

The entire WD repeats region of esc is required for binding to E(z). Many small deletions throughout this region disrupt binding (data not shown).

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

E(z) and esc were coimmunoprecipitated from a gel filtration fraction containing 600 kDa complexes.

esc, E(z), Rpd3 and Caf1 copurify in a complex with apparent molecular mass of about 600 kDa.

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

Nuclear extract was fractionated by several chromatographic steps, tracking the distribution of esc- and E(z)-containing fractions by western blot. These fractions were then subjected to anti-FLAG immunoprecipitation and component proteins identified by mass spectroscopy. Identity of copurifying proteins confirmed by western blot.

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

esc and E(z) coimmunoprecipitate from both early embryo and late embryo extracts.

Source was embryonic nuclear extract wild-type fly line; proteins produced from endogenous genes.

The complex of Su(z)12, E(z), Rpd3, esc and Caf1 has an apparent size of 600-650 kDa (gel filtration).

A histone methyltransferase activity was purified from embryonic nuclear extracts.

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

esc, E(z), Su(z)12 and Caf1 copurify as part of a polycomb repressive (PRC2) complex.

Nuclear extract was fractionated on a QAE-Sepharose comlum, then used for FLAG-affinity purification of esc containing complexes.

esc, E(z), Su(z)12 and Caf1 copurify as part of a polycomb repressive (PRC2) complex.

Source was baculovirus-Sf9 cell system; bait produced from tagged transgenic construct; prey produced from transgenic construct.

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

esc, E(z), Su(z)12 and Caf1 copurify as part of a polycomb repressive (PRC2) complex.

Nuclear extract was fractionated on a QAE-Sepharose comlum, then used for FLAG-affinity purification of esc containing complexes.

Immunoprecipitation of esc from size fractionated extracts containing complexes of about 600 kDa or 1 MDa in size.

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

esc and E(z) are present in complexes of about 600 kDa or 1 MDa. The larger 1 MDa PRC2 complex is only observed in 0-16 hr embryos, not in 18-24 hr embryos.

Purified proteins visualized by Coomassie staining.

The E(z), Caf1, Su(z)12 and esc proteins interact as part of the tetrameric Polycomb repressive complex 2 (PRC2) complex.

Analysis of direct interactions suggests that E(z), Caf1, Su(z)12 proteins form a core trimeric complex (all paiwise interactions detected) to which esc protein binds via direct interaction with E(z).

Source was extract of Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

Purified proteins visualized by Coomassie staining.

The E(z), Caf1, Su(z)12 and esc proteins interact as part of the tetrameric Polycomb repressive complex 2 (PRC2) complex.

In direct pairwise assays, tagged esc protein interacted with E(z), but not the other PRC2 complex proteins Su(z)12 or Caf1.

Analysis of direct interactions suggests that E(z), Caf1, Su(z)12 proteins form a core trimeric complex (all paiwise interactions detected) to which esc protein binds via direct interaction with E(z).

Source was extract of Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

Interaction in vitro; bait and prey produced recombinant fusion proteins in baculovirus and insect cell system.

Affinity purification carried out from a mixture of esc, E(z), Su(z)12, and Caf1-55 recombinant proteins. Deletion of E(z) II domain results in loss of Su(z)12 and Caf1-55 from the complex.

Source was cell extract of baculovirus-infected Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

Positive control.

Source was cell extract of Sf9 cell line; bait produced from baculovirus construct; prey produced from baculovirus construct.

PRC2 complexes were reconstituted from cells infected with baculoviruses for E(z), Su(z)12, Caf1-55 and esc or escl.

Su(z)12, E(z), Caf1 and esc were coexpressed in Sf9 cells and purified as a complex.

Su(z)12, E(z), Caf1 and esc form the PRC2 (Polycomb repressive complex 2).

Source was baculovirus-infected Sf9 cell extract; proteins produced from transfected constructs.

PRC2 complex of Su(z)12, E(z), esc and Caf1-55.

Source was cell extract of baculovirus-infected Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

Source was cell extract of S2R+ cell line; proteins produced from transfected construct or endogenous gene.

HGScore = 35.34381

FLAG-tagged esc was co-expressed with components of the PRC2 complex (Su(z)12, E(z) and Caf1-55) as well as Scm.

Source was cell extract of baculovirus-infected Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

The PRC2 complex consists of esc, E(z) and Caf1-55.

Interaction in vitro; bait and prey produced recombinant fusion proteins in baculovirus and insect cell system.