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. 2021 Jan 18;11:540591. doi: 10.3389/fphys.2020.540591

FIGURE 6.

FIGURE 6

miR-421 is involved in the expression of ACE2 under mechanical stretch. (A,B) RT-qPCR and Western blot analysis of ACE2 mRNA and protein expression in vascular smooth muscle cells (VSMCs) transfected with Dicer siRNA. (C) Venn diagram of the number of transcription factors in three libraries. (D) RT-qPCR analysis of the levels of miR-421 under 18% mechanical stretch for the indicated time. (E) Sequence alignment between miR-421 and putative binding sites in the 3′-UTR of ACE2 mRNA. (H) Effects of the miR-421 mimics on the activities of the luciferase reporter plasmids with the full-length (Wild) and mutated (Mut) ACE2-3′-UTR. HEK-293T cells were transfected with the ACE2-3′-UTR-WT and ACE2-3′-UTR-mut, together with miR-421 mimics, for 48 h. (F,G,I,J) RT-qPCR and Western blot analysis of ACE2 mRNA and protein expression in VSMCs transfected with miR-421 mimics (or inhibitor). (K) Western blot analysis of ACE2 protein expression in VSMCs under stretch for 12 h transfected with miR-421 inhibitor. Values are expressed as means ± SEM. *P < 0.05 vs. NC group (n = 4).