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. 2015 May 12;43(11):5524–5536. doi: 10.1093/nar/gkv470

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — RRP1 is involved in 47S/45S and 32S pre-RNA processing. (A) Total protein from HeLa cells (20 μg/lane) transfected with scRNA and empty vector (lane 1: scRNA + vector), RRP1 siRNA and empty vector (lane 2: RRP1 siRNA + vector), or RRP1 siRNA and siRNA-resistant RRP1 expression vector (lane 3: RRP1 siRNA + siResistant) was analyzed by immunoblotting (IB) with anti-RRP1 or anti-LaminB. Endogenous RRP1 and exogenously expressed FLAG-tagged siRNA-resistant RRP1 (siResistant) are indicated. (B) HeLa cells treated as above were metabolically labeled with [³H]uridine for 2 h. Total RNA was isolated and equal counts per minute (20 000 cpm) per sample were separated on an agarose gel, transferred to a membrane and detected by fluorography. Positions of rRNAs and major precursors are indicated at the left. (C) HeLa cells treated with scRNA (sc) or RRP1 siRNA (si) were labeled with [³H]uridine for 30 min, cultured in nonradioactive medium containing 0.5 mM nonradioactive uridine for the times indicated and then analyzed as in (B).