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. 2009 Apr 6;185(1):35–42. doi: 10.1083/jcb.200811106

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© 2009 Yamasaki et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Effect of endogenous angiogenin on tiRNA production and protein synthesis. (A) U2OS cells were transfected with control (D0, lanes 1 and 2), angiogenin-specific SMART pool (SP; lane 3), or angiogenin-specific individual (ANG; lane 4) siRNAs, then cultured in the absence (−; lane 1) or presence (+; lanes 2–4) of sodium arsenite (SA; 250 µM) for 1 h before Trizol extraction, separation on a 15% TBE-urea gel, and CYBR gold staining. The positions of tRNAs and tiRNAs are indicated in this representative gel. Densitometric scanning was used to quantify the relative expression of tiRNAs in three independent experiments. (B) U2OS cells were transfected with the indicated siRNAs, cultured in the absence (−; lanes 1–3) or presence (+; lanes 4–6) of sodium arsenite (100 µM, 1 h), and pulsed with [³⁵S]methionine-containing medium for 60 min before protein extraction. [³⁵S]methionine incorporation (mean ± SD, n = 3–5) is normalized to that observed in cells treated with control siRNA (designated 100%). *, P = 0.02 (n = 3); **, P = 0.04 (n = 5).