Fig. 4. Monitoring apoptosis in lamin-depleted HeLa cells. HeLa cells with depleted lamin expression were analysed (48 or 96 hours after transfection) to monitor binding of Annexin V. Cells were stained with Alexa-568 (red)-conjugated Annexin V and counterstained with Hoechst (blue) (A) and the percentage of Annexin-V-positive transfected cells (n=200) were determined (B). Cells on coverslips were treated as follows: (1) positive controls [Control (+)] treated with staurosporine (5 µm in medium; 4 hours); (2) negative controls [Control (–)] transfected with pYFP-tubulin using polyfect; (3) lamin-A/C-depleted cells (KD LA/C) transfected with pYFP-tubulin and pSECpuroLaminA/C; and (4) lamin-B1-depleted cells (KB LB1) transfected with pYFP-tubulin and pSEChygroLaminB1. Scale bars: 10 µm.