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. 1999 Jun 8;96(12):6947–6952. doi: 10.1073/pnas.96.12.6947

Figure 5.

Figure 5

Direct interaction of REA with ER. (Upper) In vitro translated, [35S]methionine-labeled REA was incubated with GST-L540Q-ER (lanes 1–3) or GST alone bound to Sepharose beads (lane 4) in the presence of 0.1% ethanol control vehicle (−), 10−6 M TOT (T), or 10−6 M estradiol (E). (Lower) In vitro translated, [35S]methionine-labeled L540Q ER (lanes 1 and 2) or wild-type ER (lanes 3 and 4) were incubated with GST-REA (REA bound to GST-Sepharose beads) in the presence of control vehicle (−) or estradiol (E). Bound protein was eluted and analyzed by 12.5% SDS/PAGE. Typically, 12–15% of input-radiolabeled wild-type ER was bound to GST-REA in the presence of ligand. The numbers at the right indicate molecular size markers in kDa.