Fig. 5. Double labelling to show concurrent -H2AX and Rad51 immunofluorescence after irradiation. Following irradiation of Namalwa cells, cell samples were taken, cytospun, fixed and stained for -H2AX and Rad51 and assessed by confocal immunofluorescent microscopy. (A) -H2AX and Rad51 foci in a Namalwa cell 4 hours after 10 Gy irradiation; red channel represents Texas Red-labelled -H2AX, green channel represents FITC-labelled Rad51 foci in the same cell. (B) Namalwa cells 3 days post irradiation. The -H2AX labelling pattern (red) has changed to confluent higher order -H2AX structures. Note the increased -H2AX immunoreactivity in smaller, presumably 4C cells (arrows) and low/absent -H2AX signal in the large Rad51-positive Namalwa endopolyploid cell (asterisks). (C) Namalwa cells, day 4 post-irradiation. The amount of Rad51 foci (green)-positive cells has increased in comparison with day 1. Clustering of the label and formation of higher-order Rad51 structures (arrow) are observed during this time period. Two adjacent or contiguous Rad51 foci was a consistent feature of endopolyploid cells (insertion). DNA was counterstained with 7-AAD. Scale bars: 20 µm.

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