American Society of Hematology

Fig. 2.

$Fig. 2. PARP cleavage induced by ceramide is prevented by elevation of cAMP. MC/9 cells grown in IL-3 were washed and resuspended in RPMI-1640 supplemented with 10% fetal calf serum. C2-ceramide (50 μmol/L) and forskolin and IBMX (10 μmol/L and 50 μmol/L, respectively) or vehicle (DMSO) were added. At the hours indicated, an aliquot of cells were harvested and lysed in sample buffer containing 6 mol/L urea, followed by sonication and boiling. Samples were fractionated by SDS-PAGE (10%) and transferred to nitrocellulose. Immunoblot analysis was performed to detect PARP (118 kD) and the corresponding 86-kD fragment. This lower fragment appeared to be cleaved further at later time points.$

PARP cleavage induced by ceramide is prevented by elevation of cAMP. MC/9 cells grown in IL-3 were washed and resuspended in RPMI-1640 supplemented with 10% fetal calf serum. C2-ceramide (50 μmol/L) and forskolin and IBMX (10 μmol/L and 50 μmol/L, respectively) or vehicle (DMSO) were added. At the hours indicated, an aliquot of cells were harvested and lysed in sample buffer containing 6 mol/L urea, followed by sonication and boiling. Samples were fractionated by SDS-PAGE (10%) and transferred to nitrocellulose. Immunoblot analysis was performed to detect PARP (118 kD) and the corresponding 86-kD fragment. This lower fragment appeared to be cleaved further at later time points.

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