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. 2015 Dec 9;44(6):2661–2676. doi: 10.1093/nar/gkv1334

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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-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 6. — NMD target stability and protein expression in Carm1 +/+ and Carm1 -/- MEFs. (A) Western blotting analysis and quantification shows levels of CARM1, UPF1 and NMD targets ASNS and ARC protein levels in Carm1 +/+ (wt) and Carm1 -/- MEFs. Tubulin was used as loading control. Data are means +/− SEM (n = 3). (B,C) Quantification of NMD transcripts Gadd45A and Asns mRNA half-lives normalized to Gapdh. Semi-logarithmic graph shows the decay rate of mRNAs calculated using formula: (x) = ln(0.5)/b, where b = slope obtained from the trendline formula: y = ne^−bx. Carm1 +/+ and Carm1 -/- MEFs were pretreated with either a vehicle or Wortmannin (5 μM) and followed by treatment with actinomycin D (5 μg/ml) and RNA extraction at 0, 2, 4, 6 and 8 h. Quantification shows the means +/− SEM (n = 3) and data are presented as relative to zero time point.