FIG. 3.

Time dependence of reconstitution of protein priming. (A) Protein-priming reactions, as described in Fig. 1, were carried out for different durations (30 min, 1 h, or 2 h), either without added chaperones or with the addition of four chaperones (Hsp90, Hsp70, Hop, and Ydj1), five chaperones (Hsp90, Hsp70, Hop, Ydj1, and p23) or reticulocyte lysate (RL). The amount of chaperones used was 120 ng for Hsp90, 350 ng for Hsp70, 125 ng for Hop, 1,000 ng for Ydj1, and 50 ng for p23. The amount of RL used was 5 μl per reaction. (B) Quantitative results derived from multiple time course experiments were summarized, and the means and standard errors are shown. The control reaction (without added chaperones or RL) signal was always set to 1, and fold activation represents the fold increase in reaction signal over this background at the respective time points. Filled circles, reconstitution with RL; open circles, reconstitution with Hsp90, Hsp70, Hop, Ydj1, and p23; open diamonds, reconstitution with Hsp90, Hsp70, Hop, and Ydj1.