American Society of Hematology

Figure 6.

$Figure 6. Caspase dependence of SAHA/bortezomib effects in K562 cells. (A) K562 cells were exposed to 1.5 μM SAHA + 4.5 nM bortezomib for 36 hours in the presence or absence 25 μM BOC-fmk, after which release of cytochrome c or Smac/DIABLO or AIF into the S-100 cytosolic fraction and the expression of procaspase-3, cleaved caspase-3, and procaspase-8 in total cell lysates were assessed as described in “Materials and methods.” (B) Cells were treated with SAHA + bortezomib ± BOC-fmk, after which Western blot analysis was used to assess expression of Raf-1, p21CIP1, cyclin D1, pRb, and Bcr-Abl. CF indicates cleavage fragment. Each lane contained 25 μg protein; blots were stripped and reprobed for tubulin to ensure equivalent loading and transfer. An additional 2 studies yielded equivalent results.$

Caspase dependence of SAHA/bortezomib effects in K562 cells. (A) K562 cells were exposed to 1.5 μM SAHA + 4.5 nM bortezomib for 36 hours in the presence or absence 25 μM BOC-fmk, after which release of cytochrome c or Smac/DIABLO or AIF into the S-100 cytosolic fraction and the expression of procaspase-3, cleaved caspase-3, and procaspase-8 in total cell lysates were assessed as described in “Materials and methods.” (B) Cells were treated with SAHA + bortezomib ± BOC-fmk, after which Western blot analysis was used to assess expression of Raf-1, p21^CIP1, cyclin D₁, pRb, and Bcr-Abl. CF indicates cleavage fragment. Each lane contained 25 μg protein; blots were stripped and reprobed for tubulin to ensure equivalent loading and transfer. An additional 2 studies yielded equivalent results.

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