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. 2019 May 16;20(10):2418. doi: 10.3390/ijms20102418

Figure 5.

Figure 5

Atg7 is a direct target of miR-27a-5p. Relative expression correlation analysis between miR-27a-5p and Atg7 during hypoxia at different timepoints (0, 3, 6, 12, 24 and 48 h after hypoxia) in H9c2 cells (A), and in AMI/sham rat visceral tissues (B). mRNA (C) and protein (D) expression of Atg7 was tested by qRT-PCR and western blotting after miR-27a-5p gain and loss of function in hypoxia-exposed H9c2 cells. (E) Schematic diagram showing the structure of dual-luciferase reporter plasmid pmirGLO and the sequence alignment of miR-27a-5p and Atg7 3′-UTR among several representative species (human, mouse and rat). Atg7 3′-UTR containing the miR-27a-5p binding site (WT or MUT) was inserted into the multiple cloning site (MSC) of pmirGLO plasmid. (F) Luciferase activity was analyzed after co-transfection of recombinant plasmid (WT or MUT) with miR-27a-5p mimic or control into HeLa cells. Three independent experiments were performed in triplicate. Data are expressed as the mean ± SD. ** p < 0.01. NC: negative control; mfe: minimum free energy.