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Individually purified wild-type and mutant USP1 is subjected to cross-linking with UAF1 using BM(PEG)2. The reaction mixtures were separated on an 8% denaturing SDS-PAGE gel and stained with Coomassie blue. No cross-linking product was observed for the wild-type (WT) USP1/UAF1, USP1(S67D)/UAF1, USP1(S67A)/UAF1, and USP1(S313A)/UAF1 complexes. A cross-linked species with a molecular weight close to 212 kDa was observed for the USP1(S313D)/UAF1 complex.
