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LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

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CHAPTER 3<br />

RESULTS<br />

Fig. 3.1.8 Response of A549 and MCF-7 cells exposed to fractionated irradiation. (A)<br />

Clonogenic Cell Survival of A549 and MCF-7 cells exposed to fractionated irradiation. Data<br />

represents means ± SE of three independent experiments; **P < 0.01 compared with MCF-7 (B)<br />

Representative image of three independent experiments showing gene expression of ATM,<br />

DNA-PK, Rad52 and MLH1 in A549 and MCF-7 cells exposed to fractionated irradiation. β-<br />

Actin gene expression in each group was used as an internal control. (C) Phosphorylation of<br />

ATM, BRCA1 and p53 in A549 and MCF-7 cells exposed to fractionated irradiation. (D) Repair<br />

kinetics as determined by γ-H2AX foci in A549 and MCF-7 cells exposed to fractionated<br />

irradiation at 15 min and 4h after irradiation. Each phospho-site antibody was indirectly labeled<br />

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

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

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

exposure time. Representative images of three independent experiments are shown here; at least<br />

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

134

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