American Society of Hematology

Figure 2.

$Figure 2. Combined treatment with adaphostin and MG-132 or bortezomib induces apoptosis in leukemia cells through induction of mitochondrial injury, caspase activation, down-regulation of Raf/MEK/ERK, and activation of JNK. Jurkat cells were treated with 400 nM adaphostin plus or minus 200 nM MG-132 or 4.0 nM bortezomib, while U937 cells were treated with 750 nM adaphostin plus or minus 250 nM MG-132, each for 8 hours. (A-B) Cytosolic (S-100) fractions were obtained as described in “Materials and methods,” and expression of cytochrome c, AIF, and Smac/DIABLO was monitored by Western blot. (C-F) At the end of the drug exposure (8 hours), cells were lysed, sonicated, and the proteins denatured and subjected to Western blot analysis using the indicated primary antibodies. For panels A and B, each lane was loaded with 20 μg protein, whereas for panels C-F, 30 μg protein was loaded in each lane. Blots were stripped and reprobed with antitubulin antibodies to ensure equal loading and transfer of protein. Results are representative of 3 separate studies.$

Combined treatment with adaphostin and MG-132 or bortezomib induces apoptosis in leukemia cells through induction of mitochondrial injury, caspase activation, down-regulation of Raf/MEK/ERK, and activation of JNK. Jurkat cells were treated with 400 nM adaphostin plus or minus 200 nM MG-132 or 4.0 nM bortezomib, while U937 cells were treated with 750 nM adaphostin plus or minus 250 nM MG-132, each for 8 hours. (A-B) Cytosolic (S-100) fractions were obtained as described in “Materials and methods,” and expression of cytochrome c, AIF, and Smac/DIABLO was monitored by Western blot. (C-F) At the end of the drug exposure (8 hours), cells were lysed, sonicated, and the proteins denatured and subjected to Western blot analysis using the indicated primary antibodies. For panels A and B, each lane was loaded with 20 μg protein, whereas for panels C-F, 30 μg protein was loaded in each lane. Blots were stripped and reprobed with antitubulin antibodies to ensure equal loading and transfer of protein. Results are representative of 3 separate studies.

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