Pre-assembled human PRC2 complex, FLAG-EED, EZH2, SUZ12 and RbAp48 proteins, was purified, from Sf9 cells co-infected with the cognate pFastBac1 constructs, by M2 anti-FLAG bead affinity column and Superose 6™ gel filtration chromatography (equilibrated with 50 m m Tris HCl pH 8.0, 150 m m NaCl, 10% glycerol).
For dual tagged complexes (FLAG-EZH2 6×HIS-EED FLAG-EZH2 6×HIS-EEDication initially FLAG-EZH2 6×HIS-EEDure for 6×His tandPRC2fication.
Silently, the crowd faced west toward the complex of flag-draped skyscrapers on the edge of the pit.
In the soluble complexes obtained, Flag and HA tagged Brk1 were still observed, arguing against a dimeric form (Fig. 3D).
We found that cyclinB, but not cyclinA, was detectable in a complex with FLAG-SIRT1 in asynchronously growing cultures (Fig. 3A).
Using immunoprecipitation assays in HEK293T cells, we detected overexpressed HA-PPARγ2 in a protein complex with FLAG-TRIM23.
In their manuscript, as a proof of principle they demonstrated the existence of the triple complex using Flag-mTOR, mCherry-Raptor and YFP-PRAS40.
We found Cdk1 in immune complexes with FLAG-SIRT1 (Fig. 3A).
Constitutively active mutants of Gαq, Gα14, and Gα16, but not their wild-type counterparts, formed complexes with Flag-Fhit as predicted.
In addition, five possible cryptic species complexes are flagged for further investigation, based on weak support for unified clustering of island populations: H. anthracinus, H. connectens, H. kukui, H. specularis, and H. volatilis.
