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. 2002 Mar;13(3):978–988. doi: 10.1091/mbc.01-05-0272

Figure 3.

Figure 3

Inhibition of PARP-1 cleavage potentiates TNF-induced death. Immortalized murine fibroblasts were triggered with TNF for 8 h. (A) Cells expressing the caspase-resistant PARP-1 D214N mutant were more sensitive to TNF-induced death than wild-type or PARP-1–deficient cells. (B) Effect of the caspase inhibitor zVAD on TNF-induced cell death in the different immortalized cells. Potentiation of TNF-induced death by zVAD was only observed in PARP-1(+/+) fibroblasts but not in PARP-1–deficient cells. SDs were <11%. (C) Expression of PARP-1 and -2 in immortalized wild-type fibroblasts and PARP knockout cells retransfected the caspase-resistant PARP D214N mutant, the vector control or wild-type (wt) PARP-1 cDNA. Cell lysates were separated by SDS polyacrylamide electrophoresis and immunoblotted with antibodies specific for PARP-1 and PARP-2, respectively. The immunoblots show the full-length form of PARP–1 (116 kDa) and -2 (62 kDa). No compensatory upregulation of PARP-2 expression was observed in the different cell clones.