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. 2022 Feb 25;163(4):bqac022. doi: 10.1210/endocr/bqac022

Figure 4.

Figure 4.

MiR-205-5p binds to 3′-UTR of GGCT and regulates the expression of GGCT. (a) Position of the miR-205-5p target site in 3′-UTR of human GGCT mRNA predicted by TargetScan (http://www.targetscan.org/). (b) Western blot analysis of the effect of miR-205-5p mimics or inhibitors on GGCT protein levels in K1 cells. (c) Schematic representation of the binding sites of miR-205-5p and the expression vectors construction of the wild type and mutant GGCT- 3′-UTR pmir-GLO plasmids used in luciferase reporter assays. (d) Luciferase reporter vectors WT-pmirGLO-GGCT (intact) or MUT-pmirGLO-GGCT (mutant) were transfected in K1 cells. The luciferase activity was measured and the values were normalized to Renilla luciferase activity. (e) Schematic of biotin-labeled RNA-RNA pull-down assays. (f) Expression of GGCT mRNA in biotin-labeled miRNA/mRNA complex was demonstrated by qRT-PCR. *: P < 0.05, **: P < 0.01.