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. 2020 Jun 18;14:1753466620915156. doi: 10.1177/1753466620915156

Figure 3.

Figure 3.

Dual-Luciferase Reporter Assay and western blotting to test binding of miR-30a-5p to, and downregulation of, target genes PIK3R2 and PIK3CD in the H1650GR cell line. (A) Putative miR-30a-5p binding sites in the 3′-UTRs of PIK3R2 and PIK3CD. A mutation was introduced into the 3′-UTR by altering 3 nt in the binding sites, and WT or Mut 3′-UTRs were subcloned into a dual-luciferase reporter vector. (B) Luciferase activities indicating binding between miR-30a-5p and PIK3R2/PIK3CD. Luc/R-luc is the ratio of firefly luciferase to Renilla-luciferase activities. *p-value < 0.05. (C) Western blotting was used to determine the PIK3R2 and PIK3CD protein expression levels in cells transfected with the miR-control or miR-30a-5p mimics. The internal control was GAPDH.

GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PIK3CD, phosphatidylinositol 3 kinase catalytic subunit delta; PI3KR2, phosphatidylinositol 3 kinase regulatory subunit 2; UTR, untranslated region; WT, wild type.