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. 2014 May 15;42(12):7981–7996. doi: 10.1093/nar/gku424

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© The Author(s) 2014. 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/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Targeted mutagenesis of suboptimal miRNA stem-loops. miR-K2 (A), miR-K5 (B) and miR-K9 (C) were mutated to optimize the secondary structure. Each missing criterion was added or changed, and the resulting mutants were cloned in the pcDNA-K10/12 or pcDNA-K2 expression plasmid. The resulting constructs were then transfected in HEK293Grip cells and total RNA was analyzed by northern blot for the studied miRNA. miR-16 was probed as a loading control. The histograms represent the relative expression of the different miRNAs from the mutated plasmids compared to the wt for three independent experiments. Inserted nucleotides are indicated in red and nucleotides derived from the miR-K6 stem-loop are indicated in blue, red crosses indicate deletions.