Fig. 3. Localization of NuMA, p85 fragment of PARP-1 and cleaved caspase-3 in cultured HeLa cells during Fas-mediated apoptosis. Cells grown on glass coverslips were treated with Fas antibody and PD 98059 for 16 hours and double-stained with NuMA (A) and the p85 fragment of PARP-1 (B) or NuMA (F) and cleaved caspase-3 (G) antibodies. FITC-conjugated goat-anti-mouse (A and F) and TRITC-conjugated goat-anti-rabbit (B and G) secondary antibodies were used. Samples were counterstained for DNA with Hoechst (D and I). In apoptotic cells, NuMA is condensed or localized outside the fragmented nuclei within the apoptotic bodies (A and F, arrows). Note that these cells are positive for cleaved p85 fragment of PARP-1 and cleaved caspase-3 (B and G). Bar, 10 µm.