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Figure 4.

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HMGB-1 protein interaction with CTE of PR is disrupted in the presence of PRE DNA. (A) Constructs of PR DBD with different lengths of CTE (aa 670, aa 651 and aa 641) were used in GST pull-down assays to detect HMGB-1 interaction. (B) Free GST or HMGB-1-GST fusion proteins were immobilized to glutathione–Sepharose resins as in Figure 1 and were incubated with PR DBD–CTE670, PR DBD–CTE651 or PR DBD–CTE641 in the absence (–DNA) or presence of PRE oligonucleotide (0.2 µM) or a nonspecific (N.S.) oligonucleotide DNA. Resins were washed, eluted and the bound protein was detected by western blotting with antibody for PR DBD. (C) GST pull-down assays with full-length PR were conducted as above in B and bound protein was detected by western blotting with a monoclonal antibody for PR.

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