Fig. 2.

sGCβ₁ subunit is a direct target of miR-34c-5p. (A) The expression of miR-34b-5p and miR-34c-5p was increased in mouse lungs treated with hypoxia. miR-34b-5p and miR-34c-5p expression was determined by quantitative real-time PCR (qRT-PCR) in the mouse lungs from the normoxic and hypoxic groups at 72 hours and 120 hours. The levels of miRNAs are relative to the control 5 s rRNA. Data are expressed as means ± s.e.m. (n = 3; *P<0.05, **P<0.01 versus control). (B) Schematic representation of the expression vector for miR-34b-5p and miR-34c-5p. (C) In vitro validation of miR-34b-5p and miR-34c-5p overexpression after transfection of HEK293T cells with pEGFP-N1-34b-5p and pEGFP-N1-34c-5p. Data are expressed as means ± s.e.m. (n = 3; ***P<0.001). (D) Wild-type putative miR-34c-5p binding site in sGCβ₁ 3′-UTR and its mutant form (top). Luciferase activity measured in HEK293T cells co-transfected with pEGFP-N1-34b-5p, pEGFP-N1-34c-5p, or empty plasmid and a luciferase reporter plasmid carrying either wild-type or mutant form of sGCβ₁ 3′-UTR (bottom). Firefly luciferase activity is normalized to Renilla luciferase expression levels. Data are expressed as means ± s.e.m. (n = 3; **P<0.01).