American Society of Hematology

Figure 6.

$Figure 6. LBH589 acetylates and inhibits hsp90 chaperone function, promoting polyubiquitylation and proteasomal degradation of Bcr-Abl and FLT-3. (A) K562 cells were exposed to the indicated concentrations of LBH589 for 8 hours. After this, hsp90 was immunoprecipitated from the cell lysates and immunoblotted with either anti-hsp90 or antiacetylated lysine antibody. (B) K562 cells were treated with the indicated concentrations of LBH589 and/or 17-AAG for 8 hours. After this, immunoprecipitates containing Bcr-Abl from the cell lysates were immunoblotted with anti-Abl or anti-hsp90 antibodies. (C) K562 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone, for 8 hours. The detergent (NP-40)-insoluble fraction was immunoblotted with Bcr-Abl antibody. The levels of β-actin served as the loading control. (D) K562 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. After this, immunoprecipitates of Bcr-Abl from the cell lysates were immunoblotted with anti-ubiquitin or anti-Bcr-Abl antibodies. (E) MV4-11 cells were exposed to the indicated concentrations of LBH589 for 8 hours. After this, hsp90 was immunoprecipitated from the cell lysates and immunoblotted with either anti-hsp90 or antiacetylated lysine antibody. (F) MV4-11 cells were treated with the indicated concentrations of LBH589 and/or 17-AAG for 8 hours. After this, immunoprecipitates of FLT-3 from the cell lysates were immunoblotted with anti-FLT-3 or anti-hsp90 antibodies. (G) MV4-11 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. The detergent (NP-40)-insoluble fraction was immunoblotted with anti-FLT-3 antibody. The levels of β-actin served as the loading control. (H) MV4-11 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. After this, immunoprecipitates of FLT-3 from the cell lysates were immunoblotted with anti-ubiquitin or anti-FLT-3 antibodies.$

LBH589 acetylates and inhibits hsp90 chaperone function, promoting polyubiquitylation and proteasomal degradation of Bcr-Abl and FLT-3. (A) K562 cells were exposed to the indicated concentrations of LBH589 for 8 hours. After this, hsp90 was immunoprecipitated from the cell lysates and immunoblotted with either anti-hsp90 or antiacetylated lysine antibody. (B) K562 cells were treated with the indicated concentrations of LBH589 and/or 17-AAG for 8 hours. After this, immunoprecipitates containing Bcr-Abl from the cell lysates were immunoblotted with anti-Abl or anti-hsp90 antibodies. (C) K562 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone, for 8 hours. The detergent (NP-40)-insoluble fraction was immunoblotted with Bcr-Abl antibody. The levels of β-actin served as the loading control. (D) K562 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. After this, immunoprecipitates of Bcr-Abl from the cell lysates were immunoblotted with anti-ubiquitin or anti-Bcr-Abl antibodies. (E) MV4-11 cells were exposed to the indicated concentrations of LBH589 for 8 hours. After this, hsp90 was immunoprecipitated from the cell lysates and immunoblotted with either anti-hsp90 or antiacetylated lysine antibody. (F) MV4-11 cells were treated with the indicated concentrations of LBH589 and/or 17-AAG for 8 hours. After this, immunoprecipitates of FLT-3 from the cell lysates were immunoblotted with anti-FLT-3 or anti-hsp90 antibodies. (G) MV4-11 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. The detergent (NP-40)-insoluble fraction was immunoblotted with anti-FLT-3 antibody. The levels of β-actin served as the loading control. (H) MV4-11 cells were either cotreated with the indicated concentrations of LBH589, 17-AAG, and PS341, and/or with the agents alone for 8 hours. After this, immunoprecipitates of FLT-3 from the cell lysates were immunoblotted with anti-ubiquitin or anti-FLT-3 antibodies.

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