Extended Data Fig. 1: Purification of a Rev1-Polζ holoenzyme complex.
a, Superdex 200 PC3.2/30 Gel filtration purification of the Rev1-Polζ holocomplex. A summary of the purification steps up to the gel filtration step are shown. 1 μl each from fractions 7-20 were separated on a 4-20% gradient SDS-PAGE gel. Molecular weight marker sizes are shown on the left and protein identities are shown on the right. b, Purified Rev1-Polζ. Fractions 8-10 from the gel filtration step (panel A) containing stoichiometric amounts of each subunit of Rev1-Polζ were pooled and incubated with 200 μl Glutathione Sepharose beads to remove any residual prescission protease and the resulting protein preparation was concentrated using a microcon MWCO-100 concentrator. Lane 1, 1.0 μl Rev1-Polζ holocomplex. Molecular weight marker sizes are shown on the left and protein identities are shown on the right. c, SDS gel profile of Rev1-ΔN-Polζ. Coomassie stained 4-20% SDS PAGE of purified Rev1-ΔN-Polζ holocomplex employed for vitrification. d, SDS gel profile of Rev1-Full-Polζ. Coomassie blue stained 4-20% SDS PAGE of purified Rev1-Full-Polζ holocomplex employed for vitrification. All the gels were run twice. Subunit identities and molecular weight markers are shown for each gel.
