Figure 2.

Binding of FAP48 to FKBP52 in Daisy T cells is prevented by FK506 and rapamycin in a dose-dependent manner. (A) Daisy cells were induced with doxycycline over 3 days, lysed, and subjected to immunoprecipitation (IP) with the anti-6-His mAb and a control mAb. Subsequently, immunoprecipitates were analyzed by Western blotting (WB) with the anti-FKBP52 pAb 173. (B) Likewise, cell lysates as in A were incubated with the anti-FKBP52 mAb EC1 or with a control mAb, and the precipitates were analyzed by Western blotting with the anti-FAP48 antiserum 766. (C) Coimmunoprecipitation experiments with the anti-FKBP52 mAb EC1 were performed on Daisy cells overexpressing FAP48. FK506 or rapamycin at the indicated concentrations was included during the incubation with Ab. Controls were performed with ethanol, which was used as vehicle. Equal loading was confirmed by Western blotting with the anti-FKBP52 pAb (Lower). Ig, immunoglobulin light chain.