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. 2019 Oct 20;11(20):8745–8759. doi: 10.18632/aging.102249

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Copyright © 2019 Ge et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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lncRNA GAS5 sponged miR-221 through both directly targeting and Ago2-dependent manner. (A) qPCR results showed that GAS5 expression level in nucleus was higher than that in the cytoplasm; (B) FISH results indicated that GAS5 was chiefly located in the cytoplasm; (C) Direct binding sites between GAS5 and miR-221 were revealed; (D) Binding relationship between GAS5 and miR-221 was detected by luciferase reporter assay with wild or mutant type of luciferase reporter plasmids of GAS5; (E) There is negative correlation between lncRNA GAS5 and miR-221 in T2D with DN tissues (N=30). (F, G) miR-221 expression was detected by qPCR. MCs were transfected with LV-GAS5 and sh-GAS5; (H, I) RIP assay was performed, which used input from cell lysate, IgG, or anti-Ago2. The expression levels of GAS5 and miR-221 were measured by qPCR.