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Fig. 2. Mechanistic differences between contagious apoptosis and spontaneous syncytial cell death. (A) Kinetic analysis of nuclear condensation observed in control syncytia or contagious apoptosis. Cells were stained as in Fig. 1A and nuclear apoptosis was measured as immunofluorescence at the indicated times after cellular fusion. (B) Effect of contagion on the syncytial size. HeLa CD4 cells were cultured overnight with healthy-(Syncytia, Co.) or PACC+-HeLa Env cells in the absence (Contagion) or presence of zVAD.fmk (Contagion+zVAD). Cells were stained as in Fig. 1A and the number of nuclei per syncytium was determined (mean ± s.d., n=4). #P<0.005. (C) Differential effect of inhibitors on spontaneous syncytial death and contagious apoptosis. Cells were primed as in B and inhibitors were added prior to co-culture (roscovitine 10 µM, purvalanol 3 µM, rapamycin 1 µM, LY294002 10 µM, pifithrin-{alpha} 10 µM and PDTC 50 µM). Nuclear pyknosis was measured as in A after 72 hours of co-culture of control syncytia, and overnight co-culture for contagious apoptosis. Results are expressed as inhibition of nuclear apoptosis as compared to untreated control cultures (mean ± s.d., n=3) #P<0.005.