FIG. 2.

HMG2 is essential for RAG-mediated coupled cleavage of IS95. Coupled cleavage reactions were performed in 2.5 mM Mg²⁺ with constant amounts of linear IS95 substrate, MBP-RAG1, and GST-RAG2, while HMG2 was added in increasing amounts in lanes 5 to 15. Lanes 1 to 4, control reactions lacking various components as indicated above the lanes; lanes 5 to 15, HMG2 at 2.5, 3.75, 5, 10, 15, 22.5, 30, 60, 90, 120, and 200 nM, respectively. (Inset) Quantitated data displayed as percent efficiency of cleavage versus HMG2 concentration (see Materials and Methods). Open symbols, cleavage at one RSS; closed symbols, cleavage at both RSSs.