Fig. 4: MiR-155 inhibits sGCβ1 expression by targeting the 3′-UTR of its transcript. | Experimental & Molecular Medicine

Fig. 4: MiR-155 inhibits sGCβ1 expression by targeting the 3′-UTR of its transcript.

From: NF-κB-responsive miR-155 induces functional impairment of vascular smooth muscle cells by downregulating soluble guanylyl cyclase

Fig. 4

a HASMCs were transfected with psiCHECK-2-sGCβ1 3′-UTR-reporter constructs [wild-type or mutant (MT)] or in combination with 80 nM of control siRNA (C), NF-κB p65 siRNA (N), control miRNA (C), miR-155 mimic (M), or miR-155 inhibitor (I), followed by stimulation with TNF-α for 24 h. Luciferase activity was determined using a dual-luciferase reporter assay kit (n = 3). b, c Transfected HASMCs were stimulated with TNF-α for 24 h. The sGCβ1 mRNA and protein levels were determined by qRT-PCR and Western blotting (n = 3). d Cells were stimulated with or without TNF-α in the presence or absence of Bay11-7082 (Bay, 5 μM) for 12 h, followed by treatment with 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (40 μg/mL) for the indicated time periods. The sGCβ1 mRNA levels were determined (n = 3). e Transfected HASMCs were stimulated with TNF-α for 24 h, followed by further incubation with SNAP (100 μM) for 24 h. The cGMP levels were determined using a cGMP assay kit (n = 6). f HASMCs were transfected with control miRNA (SMC-C) or miR-155 mimic (SMC-M), followed by co-culture with HUVECs for 24 h. The cGMP levels were determined using an ELISA kit (n = 4). *P < 0.05 and **P < 0.01

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