FIGURE 2.

Methylene blue inhibition of generation of steroid binding activity by the purified five-protein assembly system varies with the concentration of Hsp70. A, stripped GR immune pellets were incubated with Hsp90, Hop, Hsp40, and p23 in the presence of the indicated concentrations of Hsp70 and in the presence or absence of 30 μm methylene blue (MB). Immune pellets were washed, and steroid binding activity was assayed. B, a plot of the data of panel A as % inhibition of reactivation of steroid binding activity by methylene blue at each concentration of Hsp70 is shown. C, concentration dependence of methylene blue inhibition of GR reactivation in the presence of 1 (●) or 15 (○) μg of Hsp70. **, different from control at p < 0.01. D, shown is concentration dependence of methylene blue in the presence of 15 μg Hsp70 and limiting amounts of Hsp90. The upper panel presents the cpm of steroid bound, and the lower panel presents binding as a % of the control without methylene blue.