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Competition Assay. The in vitro transcription was carried out as described (19), except GAL4-FKBP was preincubated 10 min at room temperature with 10 equivalents of compound L-1 or D-1 plus 100 equivalents of competitor, prior to the addition of the reporter construct and the HeLa nuclear extract. FKBP12 was expressed in Escherichia coli as a glutathione S-transferase fusion protein and purified on glutathione agarose beads.
