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LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

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CHAPTER 3<br />

RESULTS<br />

Fig. 3.1.5 Repair kinetics as determined by γ-H2AX foci in different treatment group of A549<br />

cells at 15 min and 4h after irradiation. Cells were fixed 15 minutes and 4 hours after irradiation<br />

in 4% paraformaldehyde, permeabilized in 0.25% Triton X-100 and labeled with specific<br />

antibodies as described in “Materials and Methods”. Each phospho-site antibody was indirectly<br />

labeled with Molecular Probe 488 secondary antibody (green) and cells were mounted with<br />

ProLong Gold antifede with DAPI (blue). The encircled area roughly represents cell nuclei as<br />

seen by DAPI staining. All images were captured using Carl Zeiss confocal microscope with the<br />

same exposure time. Representative image of three independent experiments is shown here; at<br />

least 100 cells per experiment were analyzed from three independent experiments.<br />

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