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. 2004 Mar;24(5):2025–2040. doi: 10.1128/MCB.24.5.2025-2040.2004

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Copyright © 2004, American Society for Microbiology

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FIG. 8. — (A) eIF2α-S51A can collaborate in the transformation of primary human tissue. HEK cells stably transduced with simian virus 40 large T antigen and hTERT (29) were subsequently transduced with either RasV12 or eIF2α-S51A. Following selection in puromycin, two colonies per construct were examined by phase-contrast microscopy to determine morphology. (B) Expression of the indicated constructs in individual HEK clones was confirmed by Western blotting. (C) eIF2α-S51A and RasV12 confer anchorage-independent growth on HEK cells. Transformation of HEK clones expressing RasV12 or eIF2α-S51A was confirmed by plating in 0.5% soft agar for 21 days. (D) Soft agar cloning efficiency for each HEK cell line was determined by dividing the number of colonies observed by the total number of cells plated.