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. 2011 Oct 3;40(3):1065–1076. doi: 10.1093/nar/gkr805

Figure 7.

Figure 7.

Analysis of ligase activity and complex formation of a RING mutant of RAD18. (A) Purified complexes (3.7 pmol) were analyzed by SDS–PAGE followed by staining with CBB. I, RAD6A–(HisRAD18–FLAGRAD18); II, RAD6A–(HisRAD18RING–FLAGRAD18RING); III, RAD6A–(HisRAD18RING–FLAGRAD18). Structures are represented schematically, RING domains with a mutation being shown in white boxes. (B) Ligase activities of the respective RAD6A–RAD18 complexes. Increasing amounts of the complexes (0.5, 1 and 2 pmol) shown in (A) were subjected to standard assays. (C) Purified complexes (3.7 pmol) were analyzed by SDS–PAGE followed by staining with CBB. I, RAD6A–(HisRAD18ΔC1–FLAGRAD18); II, RAD6A–(HisRAD18RING–FLAGRAD18ΔC1); III, RAD6A–(HisRAD18RING/ΔC1–FLAGRAD18). (D) Ligase activities of the respective RAD6A–RAD18 complexes. Increasing amounts of the complexes (0.5, 1 and 2 pmol) shown in (C) were subjected to standard assays.