Research Paper Volume 13, Issue 11 pp 15307—15319

Constitutive activation of the NEAT1/miR-22-3p/Ltb4r1 signaling pathway in mice with myocardial injury following acute myocardial infarction

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Figure 4. NEAT1 sponging miR-22-3p reduces Ltb4r1 expression. (A) Prediction of binding sites between miR-22-3p and NEAT1 by the Lncbase v.2 website (http://carolina.imis.athena-innovation.gr/diana_tools/web/index.php?r=lncbasev2/index). (B) The expression of NEAT1 in myocardial tissues and cells of MI mice determined by RT-qPCR. (C) The verification of the binding between miR-22-3p and NEAT1 3'UTR using dual-luciferase reporter gene assay. (D) Co-localization of NEAT1 detected by FISH (× 400). (E) miR-22-3p enrichment detected by RNA-pull down. (F) Binding of NEAT1 and miR-22-3p with AGO2 tested by RIP assay. MI mice were injected with sh-NEAT1. (G) miR-22-3p expression and mRNA levels of NEAT1 and Ltb4r1 in myocardial tissues of MI mice determined using RT-qPCR. Hypoxia-induced MI cardiomyocytes were treated with sh-NEAT1. (H) miR-22-3p expression and mRNA levels of NEAT1 and Ltb4r1 in hypoxia-induced MI cardiomyocytes determined using RT-qPCR. * p < 0.05 vs. NC, Bio-probe NC, IgG, MI mice injected with sh-NC, or hypoxia-induced MI cardiomyocytes treated with sh-NC. Unpaired t-test was used to analyze data between two groups and one-way ANOVA/Tukey’s test to analyzed data among multiple groups.