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LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

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CHAPTER 3<br />

RESULTS<br />

3.4.3.4 Apoptotic DNA fragmentation in irradiated and bystander cells (EL-4 Vs EL-4)<br />

There is a clear DNA ladder formation (a hallmark of apoptosis) in irradiated and bystander EL-<br />

4 cells. But the extent of apoptosis was found to be more in irradiated cells as compared to that in<br />

bystander cells (Fig. 4, Lane 2 and 3). There was no increase in apoptosis in the cells that had<br />

received only irradiated medium (Fig. 3.4.3.4a, Lane 4) or medium from unirradiated control<br />

cells (Fig. 3.4.3.4a, Lane 5) as compared to that in unirradiated cells (Fig. 3.4.3.4a, Lane 1).<br />

Apoptosis in irradiated and bystander EL-4 cells was further confirmed by annexin V/PI assay<br />

(Fig. 3.4.3.4b).<br />

Fig. 3.4.3.4a DNA fragmentation assay of EL-4 cells. EL-4 cells cultured for 24 h in the<br />

presence of different conditioned media. DNAs of each group of cells were isolated as described<br />

in materials and methods section and electrophoresed on 1.8% Agarose gel containing ethidium<br />

bromide. Key: Lane M, DNA molecular weight marker; Lane 1, control unirradiated EL-4 cells;<br />

Lane 2, γ irradiated EL-4 cells (5Gy); Lane 3, unirradiated EL-4 cells receiving medium from γ<br />

irradiated EL-4 cells; Lane 4, unirradiated EL-4 cells receiving medium irradiated with γ rays in<br />

the absence of cells; Lane 5, unirradiated EL-4 cells receiving medium from unirradiated EL-4<br />

cells. Data shown from representative experiment of three independent experiments.<br />

177

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