Figure 4
From: IGF-1 protects tubular epithelial cells during injury via activation of ERK/MAPK signaling pathway
IGF-1 suppresses miR-429 that targets 3′-UTR of Bcl-2 mRNA to inhibit its translation in renal epithelial cells.
(A) Bioinformatics analyses for Bcl-2 targeting miRNAs showed that miR-429 had a binding site at the 3′-UTR of Bcl-2 mRNA ranged from 440th to 447th base site. (B) The levels of miR-429 in renal epithelial cells were significantly decreased In IGF-1-treated mice that received UUO. (C) We either overexpressed miR-429, or inhibited miR-429 in the purified renal epithelial cells, by transfecting the cells with plasmids carrying either a miR-429-mimic (miR-429), or a miR-429 antisense (as-miR-429). The renal epithelial cells were also transfected with a plasmid carrying null sequence as a control (null). The overexpression or inhibition of miR-429 in renal epithelial cells was confirmed by RT-qPCR. (D) MiR-429-modified renal epithelial cells were then transfected with 1 μg of Bcl-2 3′-UTR luciferase-reporter plasmid or with 1 μg of Bcl-2 3′-UTR luciferase-reporter plasmid with a site mutation between 440th to 447th base site. The luciferase activities were quantified in these cells. (E,F) Bcl-2 mRNA (E) and protein (F) in miR-429-modified renal epithelial cells. N = 10. *p < 0.05.
