Fig. 4

PELI3 is a direct target of miR-365a-5p in NSCLC cells. a The wide type (PELI3 3′UTR(WT)) and mutant (PELI3 3′UTR(MT)) miR-365a-5p targeting sequence located in the 3′UTR of PELI3 mRNA. b Luciferase reporter assay was performed in A549 and H1299 cells co-transfected with plasmid containing PELI3 3′UTR(WT) or PELI3 3′UTR(MT) and miR-365a-5p or miR-NC. c Detection of PELI3 mRNAs in biotinylated miRNA/target mRNA complex by real-time RT-PCR. The relative levels of PELI3 mRNA in the complex pulled down by using biotinylated miR-365a-5p was compared to that of the complex pulled down by using the biotinylated control random RNA. d–f The relative expression levels of PELI3 in A549 and H1299 cells transfected with indicated microRNA mimics and microRNA inhibitors or their respective negative controls detected by qRT-PCR and western blot. g A statistically inverse correlation between PELI3 and miR-365a-5p levels in 40 NSCLC tissues (Spearman’s correlation analysis). The data represent the mean ± SD from three independent experiments. *P < 0.05; **P < 0.01