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Figure 3 | BMC Genomics

Figure 3

From: A highly expressed miR-101 isomiR is a functional silencing small RNA

Figure 3

miR-101 and 5’-isomiR-101 differentially regulate gene expression. A. mRNA expression of miR-101 target genes following exogenous expression of miR-101 or 5’isomiR-101. After transfection of SH-SY5Y cells with the siGLO control sequence (Con) or the mIRIDIAN mimics for miR-101 or 5’-isomiR-101 the expression of COX-2, MCL-1, APP, EZH2 and DUSP-1 (MKP-1) was determined by RT-qPCRs. A control sample was used to normalize Fold Change values in each set of experiments. This control sample was assigned a value of 1. B. Protein expression of miR-101 target genes following exogenous expression of miR-101 or 5’isomiR-101. Western blot determination of COX-2, MCL-1, APP, EZH2 and DUSP-1 (MKP-1) levels in SH-SY5Y were performed in cells transfected with a control sequence (Con) and the mIRIDIAN mimics for miR-101 or 5’-isomiR-101. A control sample was used to normalize Fold Change values in each set of experiments. This control sample was assigned a value of 1. C. Expression of miR-101 target genes cells transfected with miR-101 and 5’-isomiR-101 biotynilated isomiRs. SH-SY5Y cells were transfected with Biotin, mIRIDIAN 3’-biotin-miR-101 or 3’-biotin-5’-isomiR-101. Biotin-miRNA complexes were pulled down with streptavidin beads and the expression of COX-2, MCL-1, APP, EZH2 and DUSP-1 (MKP-1) mRNA was determined by RT-qPCRs. Results are expressed as the Fold Change ratios obtained from the biotin-tagged miR-101 and 5’-isomiR-101 transfected cells versus the biotin transfected control cells. ND: Not detectable. All data are expressed as the mean ± SEM from at least three independent experiments. Asterisks indicate statistical significance between miR-101 and 5’isomiR-101 data: ** (p≤0.01), * (p≤0.5) using the Mann–Whitney test.

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