Fig. 3

Hypoxia down-regulates EPDR1 expression in BC cells through up-regulation of miR-181a-5p. (A) The differentially expressed genes in the GSE49999 dataset were analyzed by GEO2R online analysis software. (B) The potential binding site was between miR-181a-5p and EPDR1 mRNA. (C) The expression of miR-181a-5p in BC was analyzed by ENCORI. (D) The relation between miR-181a-5p and BC prognosis was analyzed by Kaplan-Meier Plotter. (E) The binding of miR-181a-5p to EPDR1 mRNA was confirmed by dual luciferase reporter gene assays; *, vs. mimics NC group, p < 0.05. (F) The binding of miR-181a-5p to EPDR1 mRNA was confirmed by AGO2-RIP; *, vs. IgG group, p < 0.05. Transfection of miR-181a-5p mimics and inhibitor for BC cells, (G) the expression of miR-181a-5p and EPDR1 mRNA was examined by RT-qPCR; (H) the expression of EPDR1 protein was examined by western blot assay; *, vs. mimics NC group, p < 0.05; #, vs. inhibitor NC group, p < 0.05. Transfected hypoxia-treated MCF-7 cells with miR-181a-5p inhibitor, (I) the expression of miR-181a-5p and EPDR1 mRNA was examined by RT-qPCR; (J) the expression of EPDR1 protein was examined by western blot assay; *, vs. Normoxia group, p < 0.05; #, vs. Hypoxia group, p < 0.05; $, vs. Hypoxia + inhibitor NC group, p < 0.05