Fig. 7.
Fig. 7. KF58333 treatment destabilizes Hsp90/p210 Bcr-Abl complex. / The effects of KF58333 on Hsp90/p210Bcr-Abl complex (A and B) and the half-life of Bcr-Abl protein (C) are shown. (A) K562 cells were treated with 0.05 μmol/L of KF58333 for 3 hours, total lysate was prepared, resolved by SDS-PAGE, and analyzed by Western blotting as described previously. (B) About 1 mg of total cell lysate was immunoprecipitated with anti-p210Bcr-Abl monoclonal antibody and analyzed by Western blotting as described. (C) The half-life of Bcr-Abl protein in K562 cells treated with or without KF58333 (0.05 μmol/L) was determined as described in “Materials and methods.” 35S-methionine–labeled Bcr-Abl protein was immunoprecipitated and resolved by SDS-PAGE. Gels were fixed, enhanced, and exposed to Kodak XAR-5 film. (D) Bcr-Abl–specific bands were quantified by densitometric scanning/image analysis. 35S,35S-methionine; ip, immunoprecipitated.

KF58333 treatment destabilizes Hsp90/p210 Bcr-Abl complex.

The effects of KF58333 on Hsp90/p210Bcr-Abl complex (A and B) and the half-life of Bcr-Abl protein (C) are shown. (A) K562 cells were treated with 0.05 μmol/L of KF58333 for 3 hours, total lysate was prepared, resolved by SDS-PAGE, and analyzed by Western blotting as described previously. (B) About 1 mg of total cell lysate was immunoprecipitated with anti-p210Bcr-Abl monoclonal antibody and analyzed by Western blotting as described. (C) The half-life of Bcr-Abl protein in K562 cells treated with or without KF58333 (0.05 μmol/L) was determined as described in “Materials and methods.” 35S-methionine–labeled Bcr-Abl protein was immunoprecipitated and resolved by SDS-PAGE. Gels were fixed, enhanced, and exposed to Kodak XAR-5 film. (D) Bcr-Abl–specific bands were quantified by densitometric scanning/image analysis. 35S,35S-methionine; ip, immunoprecipitated.

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