Fig. 6

CircNFIB1 antagonizes miR-486-5p-mediated repression of PIK3R1 expression. a-d Representative images and a histogram analysis of tube formation and Transwell assay by HLECs treated with conditioned medium from miR-486-5p-silenced (a and b), miR-486-5p overexpressing (c and d), and corresponding control PDAC cells. Scale bar: 100 μm. e Schematic illustration showed the alignment of miR-486-5p with PIK3R1 and the red portion indicated the mutagenesis nucleotides. f Dual luciferase reporter assays showed the luciferase activity of wild type or mutant PIK3R1 following co-transfection with miR-486-5p mimic or control mimic. Relative firefly luciferase expression was normalized to that of Renilla luciferase. g and h The effect of circNFIB1-silencing or circNFIB1 overexpression on PIK3R1 expression in PDAC cells was assessed by qRT-PCR. i and j qRT-PCR analyzed the effect of circNFIB1-silencing on miR-486-5p depletion-induced PIK3R1 expression in PDAC cells. k and l Western blot analysis of circNFIB1-silencing on miR-486-5p depletion-induced PIK3R1 expression in PDAC cells. Significance level was assessed using two-tailed Student t-tests and one-way ANOVA followed by Dunnett’s tests for multiple comparisons. Figures with error bars showed the standard deviations of three independent experiments. *p < 0.05 and **p < 0.01