Neuronally differentiated PC12 cells are rescued from NGF deprivation, but not from SOD1 downregulation, by V-ANedd.A, V-ANedd differentially protects neuronally differentiated PC12 cells from apoptosis caused by NGF withdrawal. PC12 cells were neuronally differentiated by exposure to NGF (100 ng/ml) for at least 7 d in RPMI 1640 medium plus 1% horse serum. Cells were deprived of serum and NGF and replated as described in Figure 4. Additives present at the time of plating included 400 nmV-ANedd, 400 nmV-ICE_inh, or 100 ng/mlNGF (4 left-hand bars). For SOD1 downregulation (4 right-hand bars) neuronally differentiated PC12 cells were plated in RPMI 1640 medium plus 1% horse serum, with 100 ng/ml NGF. At the time of plating, cultures were incubated, as indicated, with 50 nmV-ASOD1and with the indicated additives (800 nmV-ANedd and 50 nmV-ICE_inh). Cell survival was determined after 1 d and expressed as in Figure 4.B, Dose–response curve for protection from NGF deprivation by V-ANedd. Neuronally differentiated PC12 cells were washed as above for NGF deprivation and plated in serum-free medium with the indicated concentrations of V-ANedd. Cell survival relative to the number present with the addition of NGF was quantified at 1 d.C, V-ANedd provides long-term protection against NGF deprivation. Neuronally differentiated PC12 cells were deprived of NGF and serum and plated as described in A. V-ANedd (400 nm) was included at the time of NGF deprivation and replenished 1 d later. Cell survival was determined at the indicated times as in Figure 4.