Figure 5

The effect of bps6197 mutations is reconstituted using oligonucleotide substrates. (A) A radiolabeled 23-RSS substrate, a wild-type bps6197 oligonucleotide substrate, or bps6197 substrates bearing a 23-RSS coding flank substitution (bps6197 SE/23RSS CE) or mutations in the inverted repeat (bps6197 mIR) or nonamer (bps6197 mNon) sequences were incubated in an in vitro cleavage reaction containing WT cMR1/cMR2 with or without added HMGB1 and cold partner 12-RSS (1.0 pmol) as indicated. Reaction products were analyzed as in Figure 3A. (B) WT cMR1/cMR2 was incubated with the radiolabeled substrates in (A) in binding reactions lacking or containing HMGB1. Protein-DNA complex formation was analyzed by EMSA as in Figure 3B.