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Effect of eEF-2K down-regulation on breast cancer cell proliferation and clonogenicity.

By Ibrahim Tekedereli (149655), S. Neslihan Alpay (149661), Clint D. J. Tavares (149668), Zehra E. Cobanoglu (149674), Tamer S. Kaoud (149681), Ibrahim Sahin (149690), Anil K. Sood (149701), Gabriel Lopez-Berestein (149712), Kevin N. Dalby (149722) and Bulent Ozpolat (149735)

Abstract

<p>(<b>A</b>) Knockdown of eEF-2K by siRNA. siRNA specifically targeting eEF-2K inhibits its expression in MDA-MB-231 cells. Cells were transfected with eEF-2K siRNA (5–75 nM) for 48 h, and cell lysates were subjected to Western blot analysis. (<b>B</b>) Effect of knockdown of eEF-2K on cell proliferation. MDA-MB-231 cells were transfected with eEF-2K siRNA, and after 48 h proliferation was detected by an MTS assay. (<b>C</b>) Overexpression of eEF-2K in MDA-MB-231 cells. Cells were transiently transfected with an expression vector encoding GST-tagged wild type eEF-2K, and cell lysates (48 h) were subjected to Western blot analysis. For the effect of overexpression of eEF-2K on cell proliferation, MDA-MB-231 cells were transiently transfected with eEF-2K or a vector control, and cell proliferation was measured using an MTS assay after 96 h. (<b>D–E</b>) Effect of eEF-2K knockdown on colony formation. Knockdown of eEF-2K by siRNA (50 nM) significantly inhibited the number of colonies formed by MDA-MB-231 (D) and MCF-7 (E) cells (*p<0.05). Cells were transfected every 4 days with control or eEF-2K siRNA.</p

Topics: Genetics, Molecular Biology, Cancer, eef-2k, down-regulation, cancer, proliferation
Year: 2012
DOI identifier: 10.1371/journal.pone.0041171.g001
OAI identifier: oai:figshare.com:article/276213
Provided by: FigShare
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