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Fig. 5. Contagious apoptosis is associated with caspase-resistant foci of DNA lesions. (A) DNA fragmentation occurring during contagious apoptosis. Cells and syncytia generated as in Fig. 1 were subjected to simultaneous Hoechst 33342 and TUNEL staining. Note that only cells with full-blown chromatin condensation (but not those with PACC) are TUNEL-positive. (B) Representative images of cells stained with antibodies specifically recognizing phosphorylated H2AX (Ser 139, H2AXP) or Chk2 (Thr68, Chk2P) and counterstained with Hoechst 33342. (C,D) Transmission of H2AX and Chk2 phosphorylation to nuclei from HeLa CD4 cells. Prior to co-culture, healthy HeLa CD4 were stained with CellTracker® Red. After fusion with either untreated HeLa Env cells (Co.), STS-pulsed HeLa Env cells in the absence (Contagion), or presence of Z-VAD.fmk, the resulting syncytia were fixed, permeabilized and stained for the detection of Chk2P (C) or H2AXP (D), revealed with an anti-IgG Alexa® Fluor Green conjugate. Nuclei were counterstained with Hoechst 33342. This experiment was repeated twice, with similar results.
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