pone.0017123.g001.tif (1.87 MB)

Effect of ionizing irradiation either alone or in combination with pUC and pU2 in human meningioma cell lines.

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posted on 11.02.2011, 00:26 by Reshu Gupta, Arun Kumar Nalla, Venkateswara Rao Gogineni, Chandramu Chetty, Praveen Bhoopathi, Jeffrey D. Klopfenstein, Andrew J. Tsung, Sanjeeva Mohanam, Jasti S. Rao

(A, B) Conditioned medium of two meningioma cell lines, IOMM-Lee and CH-157MN, was collected after giving 5 and 10 gy doses of γ-rays irradiation. Human microvascular endothelial cells were seeded in 96-well plates and cultured with conditioned medium of meningioma cells. The cells were maintained for 48 hrs at 21% O2, washed, fixed, stained with Hema-3, and photographed. The percentages of angiogenic cells were assessed using Cell Quest software. Columns represent means ± SD of three independent experiments. *p<0.05, significant difference from pSV (C) 1×105 IOMM-Lee and CH-157MN cells were transfected with pSV, pUC and pU2 and cell lysates were subjected to SDS-PAGE, and immunoblot analysis was carried out to detect uPAR level. Similarly, tissue lysates isolated from nude mice were also subjected for western blotting to detect uPAR expression. (D) Human microvascular endothelial cells were seeded in 48-well plates and cultured with conditioned medium collected from pSV, pUC and pU2 transfected IOMM-Lee and CH-157MN meningioma cells. The cells were kept in a humidified atmosphere containing 5% CO2 at 37°C for 48 h, washed, fixed and stained with Hema-3 and photographed. (E) Columns represent means ± SD of three independent experiments. * and # denotes p<0.05, significant difference from pSV and pSV+IR, respectively.

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