American Society of Hematology

Figure 7

$Figure 7. PP2-mediated apoptosis is countered by specific procasp-8 inhibition. (A) Freshly isolated B-CLL cells were transfected by nucleofection with either the control-siRNA (lanes 1-8) or procasp8-siRNAs (9-16), cultured for 48 hours in complete medium, and subsequently incubated in the absence (lanes 1-4 and 9-12) or presence (lanes 5-8 and 13-16) of 10 µM PP2 at different time points. After such treatment, cells were lysed and analyzed by Wb analysis with anti-procasp8, anti caspase-3, and anti-PARP antibodies as well as anti-β-actin antibody as a loading control. (B) Freshly isolated B-CLL cells were cultured as described in A. After treatment with the aforementioned inhibitor, cells underwent differential centrifugation to separate mitochondrial and cytosolic fractions. Equivalent protein amounts were assayed by Wb analysis with anti-cyt-c antibody as well as with anti-aconitase and anti-LDH antibodies to assess the purity of the fractions.$

PP2-mediated apoptosis is countered by specific procasp-8 inhibition. (A) Freshly isolated B-CLL cells were transfected by nucleofection with either the control-siRNA (lanes 1-8) or procasp8-siRNAs (9-16), cultured for 48 hours in complete medium, and subsequently incubated in the absence (lanes 1-4 and 9-12) or presence (lanes 5-8 and 13-16) of 10 µM PP2 at different time points. After such treatment, cells were lysed and analyzed by Wb analysis with anti-procasp8, anti caspase-3, and anti-PARP antibodies as well as anti-β-actin antibody as a loading control. (B) Freshly isolated B-CLL cells were cultured as described in A. After treatment with the aforementioned inhibitor, cells underwent differential centrifugation to separate mitochondrial and cytosolic fractions. Equivalent protein amounts were assayed by Wb analysis with anti-cyt-c antibody as well as with anti-aconitase and anti-LDH antibodies to assess the purity of the fractions.

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