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. 2011 Aug 25;7(8):e1002194. doi: 10.1371/journal.ppat.1002194

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Zhou et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Pull down assays with high-speed cytosolic extracts (HSE) and GST-Tnp3 in the presence or absence of RanQ69L-GTP. Proteins bound to the beads were eluted and analyzed by 15% denaturing SDS-PAGE and silver staining. M, molecular weight protein markers; lane 1, beads only + HSE; lane 2, Tnp3 + HSE + RanQ69L-GTP; lane 3, Tnp3 + HSE without RanQ69L-GTP. The arrow points to the ∼20 kDa tRNA band. (B) Nucleic acids were purified from the same fractions, analyzed by 15% denaturing PAGE and visualized following staining with SYBRgold. Lane M, 10 bp ladder; Lane 1, total RNA from 293T cells; Lane 2, small RNA fraction from 293T cells; Lane 3, in vitro synthesized control tRNA; Lane 4, same tRNA x3; Lane 5, HSE; Lane 6, Tnp3 + HSE + RanQ69LGTP; Lane 7, Tnp3 + HSE without RanQ69LGTP.