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. 2020 Feb 10;17(5):651–662. doi: 10.1080/15476286.2020.1722450

Figure 3.

Figure 3.

miR-124 and miR-128 regulate TREK1 and TREK2 expression. (A, B). SK-N-SH cells were co-transfected with Trek1 or Trek2 CDS plus 3ʹUTR plasmids and either pri-miRNA expression plasmids or 2ʹ-O-methyl miRNA inhibitors (antagomiRs). 48 hours later, Trek1 and Trek2 mRNA levels were measured by RT-qPCR. Gapdh mRNA was used as an internal reference control. These assays demonstrated that miR-124 overexpression significantly reduced Trek1 and Trek2 mRNA levels, while anti-miR-128 significantly increased Trek1 and Trek2 mRNA levels, compared to scramble controls. Trek2 mRNA levels were also increased by anti-miR-124 and anti-miR-183. (C, D). SK-N-SH cells were co-transfected with Trek1 or Trek2 CDS+3ʹUTR plasmids and either pri-miRNA expression plasmids or 2ʹ-O-methyl miRNA inhibitors. 48 hours later, TREK1 and TREK2 protein levels were measured by immunoblotting. GAPDH was used as an internal control for loading. These assays demonstrated that miR-124 and miR-128 overexpression significantly reduced TREK1 and TREK2 levels, while TREK2 protein levels were also reduced by miR-183 overexpression. Anti-miR −124 and −128 oligos significantly increased TREK1 levels, compared to scramble controls, while TREK2 levels were increased by all three anti-miR oligos. Data show the mean ± SEM from 4 RT-PCR and 5 immunoblotting independent experiments (*, p < 0.05; **, p < 0.01; ***, p < 0.001).