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. 2011 May 19;39(16):7179–7193. doi: 10.1093/nar/gkr370

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Involvement of TRF4-2 in PROMPT 3′-end adenylation. (A) Decrease in transcript levels upon RNAi-depletion was measured with gene-specific primers by RT–qPCR. Levels were normalized to those of a control KD with siRNAs directed against eGFP and to levels of GAPDH mRNA. Error bars show standard deviations of duplicate technical replica of a representative experiment. (B) Western blotting analysis of RRP40 levels in siRNA-mediated KDs as indicated above each lane. HeLa whole-cell protein extracts were separated on a 10% polyacrylanide gel, blotted to membrane and probed with an anti-RRP40 antibody (top panel). Actin levels served as a loading control (bottom panel). Migration and size of marker proteins is given to the left. (C) Effects on PROMPT levels of KD's of nuclear poly(A) polymerases alone or in combination with an RRP40 KD. RTqPCR was performed as in (A). Fold differences for three representative PROMPTs, as compared to a control KD, are shown on the y-axes.