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Native Polyacrylamide Gel Electrophoresis Immunoblot Analysis of Endogenous IRF5 Dimerization
Published on: October 6, 2019
miR-148-1-5p modulates NF-κB signaling pathway by targeting IRAK1 in miiuy croaker (Miichthys miiuy)
Renjie Chang1, Weiwei Zheng1, Qiang Luo1
1Laboratory of Fish Molecular Immunology, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China.
Abstract:
microRNAs (miRNAs), a crucial class of small non-coding RNA species, have been extensively studied as key molecular in immune regulation in the past decades. Here, we discover a new miRNA miR-148-1-5p and we elaborate that miR-148-1-5p functions as a negative regulator to participate in innate immune responses. In this article, it has been researched that the regulation effect of miR-148-1-5p to the nuclear factor kappaB (NF-κB) signaling pathway by targeting IRAK1 in miiuy croaker. First, through bioinformatics software to predict the potential targets of miR-148-1-5p, we found that IRAK1 had a base complementary region with indicated miRNA. Next, the dual-luciferase assays revealed that overexpression of miR-148-1-5p mimics and pre-miR-148 plasmid could significantly inhibit the luciferase activity of wild-type IRAK1-3'UTR. However, miR-148-1-5p inhibitors attenuated the inhibition caused by miR-148-1-5p. In addition, we also confirmed that miR-148-1-5p could suppress the expression of IRAK1 at mRNA level. Collectively, the regulations of miR-148-1-5p to NF-κB signaling pathways via targeting the IRAK1 gene was studied in miiuy croaker, which provided new information to enrich the immune regulation network of miRNA in teleost fish.
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