Skip to main content
. 2008 Feb 14;36(6):2060–2072. doi: 10.1093/nar/gkn049

Figure 2.

Figure 2.

The RAG1 NTD is necessary, but not sufficient, for stabilizing association of Ku70/Ku80 to a RAG–RSS complex. (A) Anti-Ku70 and anti-Ku80 antibodies supershift FLMR1/cMR2 RAG–RSS complexes. Radiolabeled intact 12-RSS substrate was incubated with purified Ku70/Ku80, or with HMGB1 and the various RAG preparations purified using the mild protocol, and subjected to supershift analysis by EMSA using purified monoclonal antibodies to Ku70, Ku80 or MBP, as indicated above the gel. Protein-DNA complexes supershifted by anti-Ku antibodies are indicated by arrows with an asterisk at right. (B) Supplementing RAG-RSS binding reactions with purified Ku70/Ku80 fails to supershift RAG–RSS complexes. Radiolabeled intact 12-RSS substrate was incubated with the various RAG preparations shown in Figure 3B in binding reactions containing HMGB1 in the absence or presence of purified Ku70/Ku80 as indicated above the gel, and then RAG–RSS complex formation was analyzed by EMSA.