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. 2016 May 19;11(5):e0156065. doi: 10.1371/journal.pone.0156065

Fig 3. The miR-590 C57T SNP reduces levels of miR-590-5p and miR-590-3p, without affecting the levels of pri-miR-590 and pre-miR-590.

Fig 3

(A) Quantification of pri-miR-590 by qRT-PCR normalized by GAPDH. Mean ± SD (n = 3). HEK293T cells were transfected with the pri-miR-590-WT or pri-miR-590-SNP plasmids. The empty plasmid was used as negative control. (B-F) Northern blot images for pre-miRNA, miRNA, and U6 RNA using total RNA prepared from HEK293T cells transfected with the pri-miR-590-WT or pri-miR-590-SNP plasmids. The empty plasmid was used as negative control. Four biological replicates were analyzed for each transfection plasmid. Northern probes used are perfectly complementary to miR-590-5p (A), miR-590-3p-WT (B), miR-590-3p-SNP (C), miR-16-5p (D), and U6 RNA (E). The miR-590-3p-WT probe weakly cross-hybridized to miR-590-3p-SNP, and vice versa. (G) The abundance of pre-miR-590, miR-590-5p and miR-590-3p-(WT/SNP) relative to the mean value of miR-590-5p in the WT miR-590 gene plasmid transfection conditions. (H) The abundance of miR-16-5p normalized to the mean value of the pri-miR-590-WT plasmid transfection conditions. Mean ± SD (n = 4).