Fig. 5

circAKT3 exerts its function by sponging miR-198. a & b Schematic illustration showing the overlap of the target miRNAs of circAKT3 predicted by miRanda, PITA and RNAhybrid. c & d Lysates prepared from SGC7901CDDP and BGC823CDDP cells stably transfected with circAKT3 or vector were subjected to RNA pull-down and tested by RT-PCR (C) and RT-qPCR (D). The relative level of circAKT3 was normalized to the input. GAPDH served as a negative control. e & f The relative levels of 11 miRNA candidates in SGC7901CDDP and BGC823CDDP lysates were detected by RT-qPCR. Multiple miRNAs were pulled down by circAKT3, and miR-198 was pulled down by circAKT3 in both cell lines. g Schematic illustration showing the 3′UTR of luciferase reporters containing the complete circAKT3 sequence (luc-wt) or the circAKT3 sequence with deletions of miR-198 (luc-m1-m8) binding sites. h Reporter assays showing the luciferase activity of luc-wt and luc-m1-m8 in 293 T cells cotransfected with miR-198 mimics or a scrambled oligonucleotide (control). i FISH showing the colocalization of circAKT3 and miR-198 in SGC7901CDDP cells. Nuclei were stained with DAPI. Scale bar, 10 μm. The results are presented as the mean ± SEM. *P < 0.05, **P < 0.01