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LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

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CHAPTER 3<br />

RESULTS<br />

Fig. 3.1.7 Translocation of phospho-p53 into the nucleus of A549 cells 18h after irradiation. (A)<br />

Representative image of three independent experiments showing phosphorylation and<br />

translocation of p53 into nucleus of A549 cells. 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). All images were captured using Carl Zeiss confocal<br />

microscope with the same exposure time. At least 100 cells per experiment were analyzed from<br />

three independent experiments. (B) Representative immunoblot image of three independent<br />

experiments depicting phosphorylation of p53 in different treatment group. Graph represents<br />

phospho-p53 band intensities (divided with the respective loading control intensities and<br />

normalized). Key Lane 1, control unirradiated A549 cell line. Lane 2, 2 Gy acute dose (single<br />

dose that was delivered in each fraction) of 60 Co γ-irradiation Lane 3 2 Gy of 60 Co γ-irradiation<br />

daily over a period of 5 days, Lane 4, 10 Gy acute (which is the sum total of all the fraction)<br />

60 Co γ-irradiation.<br />

133

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