35S-methionine-labeled Ub-GFP, Ub(K48R)-GFP, and Ub(K11R)-GFP translated in the PURE system were incubated with E1, E2 (UBE2A), His-Ub, ATP, and recombinant UBR4 and KCMF1. The samples were then analyzed by autoradiography either directly (input) or after His-Ub PD. See alsoFigure S9.
