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. 2013 Nov 20;3:3277. doi: 10.1038/srep03277

Figure 3. Carboplatin- and cisplatin-induced DNA damage responses and apoptosis induction in cells lacking and expressing DNA polymerase η.

Figure 3

(a) Polη-deficient XP30RO cells and polη-expressing TR30-2 cells were treated with carboplatin or cisplatin and harvested 24 h post-treatment. Polη, DNA damage response proteins and apoptosis-related proteins were analysed by western blotting using specific antibodies, as described in Materials and Methods. For direct comparison, samples from both cell lines were run on the same gel. As a control for apoptosis induction, proteins, obtained from floating cells following treatment of cells with 5 μg/ml cisplatin for 24 h, were loaded in the lanes marked A. (b) Polη-deficient XP30RO cells were treated for 24 h with 8.5 μM cisplatin. Immunofluorescence microscopy was carried out using phospho-specific primary antibodies against γH2AX and phospho-Ser4/Ser8 RPA2, and Alexa 594- and Alexa 488-labeled secondary antibodies. DNA was counterstained with DAPI. Scale bar corresponds to 25μm.