(A) Size exclusion chromatography of His–MBP–Pch2 purified from insect cells. Coomassie Brilliant Blue (CBB) staining of peak fractions (dotted line) run on SDS–PAGE gel. * indicates likely monomeric fraction of His–MBP–Pch2. AU, arbitrary units. (B, C) Amylose-based pulldown of the ORC (His–Orc1-6 and His–ORC) purified from insect cells, with His–MBP–Pch2. (B) CBB staining, (C) Western blot analysis using α-MBP and α-ORC (which recognizes all six ORC subunits). (D) Size exclusion chromatography of His–ORC (His–MBP–Pch2) assembly. CBB staining of peak fractions (dotted line) run on SDS–PAGE gel. AU, arbitrary units.