Fig. 4

CircEMB sponges miR-3184-5p. a-bRIP assay was performed to verify the binding between circEMB and the Ago2 protein. c Schematic representation of the overlap among target miRNAs of circEMB predicted using the TargetScan, miRanda, RNAhybrid, ENCORI and RNA22 databases. d-e RNA pull-down assay verified the binding between seven candidate miRNAs and circEMB. f-h RIP assay of miRNA showed that in contrast to the NC group, the miR-3184-5p mimics group effectively suppressed circEMB levels in 143B and U2OS cells. i-j HEK293T cells were co-transfected with miR-3184-5p mimics and wild-type or mutant circEMB luciferase reporter vector, and luciferase reporter activity was assessed. k Subcellular colocalization between circEMB (red) and miR-3184-5p (green) was observed in in 143B and U2OS by confocal FISH. Scale bar, 50 μm. Data are expressed as mean ± SD (n = 3) (*, p < 0.05; **, p < 0.01; ***, p < 0.001)