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Figure 3


Fig. 3. Optimisation of treatment with DNA-damaging agents. (A) Western blot analysis of untransfected HeLa cells and cells transfected with wild-type GFP-lamin A, treated with cisplatin (CP) for 0-24 hours, or UV irradiation with a recovery period of 0.5 or 24 hours, and probed with antibodies to PARP-1 and lamin A/C. Arrowheads indicate cleavage products as a result of apoptosis. Wild-type GFP-lamin A is indicated by an asterisk. Molecular mass markers indicated on the left are: phosphorylase b, 94 kDa; albumin, 67 kDa; ovalbumin, 43 kDa; and carbonic anhydrase, 30 kDa. (B) HeLa cells treated with cisplatin for 4 hours or UV irradiation with a recovery time of 0.5 hours were stained with antibody to lamin A/C (LA-2B3). (C) HeLa cells treated with cisplatin for 0-24 hours were stained with antibody to {gamma}-H2AX, and counterstained with DAPI. Arrowheads indicate fragmented nuclei. Percentages of apoptotic cells after cisplatin treatment were <3% (4h), 5% (8h), 15% (16h) and 40% (24h); and after UV irradiation, <3% (0.5h) and 50% (24h). Bar, 10 µm.





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