TPR cofactors differentially recruit components of the multichaperone heterocomplex.

<p>HEK cells were transfected and TPR cofactors immunoprecipitated as described in the legends to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011717#pone-0011717-g007" target="_blank">figures 7</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011717#pone-0011717-g008" target="_blank"></a><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011717#pone-0011717-g009" target="_blank"></a><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011717#pone-0011717-g010" target="_blank">10</a>. The relative amounts of the precipitated TPR cofactors, the co-precipitated Hsp70 and Hsp90 were determined by densitometry of a coomassie stained gel of the eluates (A, upper panel), and the relative amount of p23 by densitometry of the immunoblot signals (A, lower panel). B and D, quantification of the relative binding of co-precipitated Hsp90 (B) and Hsp70 (D). Hsp90 signals and Hsp70 signals (only intensities above background binding were taken into consideration) were normalized to signals of the respective precipitated TPR cofactors. Data are presented as relative binding + S.E.M. of at least 12 independent experiments with different steroid receptors. C, quantification of the relative binding of co-precipitated p23. The p23 immunoblot signals were related (normalized) to the respective TPR cofactor signal. Binding of p23 is presented relative to the mean of the normalized p23 eluate signals of the complete set of TPR-proteins.