Figure EV2. Linkage specificity of RNF114‐mediated polyubiquitin chains.

• A, B
  Ubiquitin mutants (K63R, K48R, K33R, K29R, K27R, K11R, K6R, or K11 only, K6 only, K27 only, K29 only, K33 only, K48 only, K63 only, and K0) were used for ubiquitin chain assembly. K0 indicates that all seven internal lysine residues were mutated to arginine. In the absence of K6, K29, K33, or K63 residues (K6R, K29R, K33R, or K63R), the mutated ubiquitins could still be used to assemble polyubiquitin chains, while in the absence of K11, K27, or K48 (K11R, K27R, or K48R), the mutated ubiquitins could not be used to assemble polyubiquitin chains by RNF114. K11‐only or K27‐only ubiquitin could be used to assemble polyubiquitin chains by RNF114.