Fig. 4

Hsa_circ_0026416 acts as a ceRNA for miR-346. a A schematic illustration showing the predicted binding sites of miRNAs associated with hsa_circ_0026416. b Venn diagram for overlaps between the two databases (RegRNA2.0 and CircInteractome). c Relative luciferase activity of candidate miRNAs after co-transfection of miRNA mimics and pmirGLO-circ_0026416 in HEK293T cells. d Expression of miR-346 in CRC tissues and ANM from TCGA data. e qRT-PCR for expression of miR-346 in 24 pairs of CRC and normal colorectal tissues. f qRT-PCR for expression of miR-346 in HCT-8 or SW480 cells after transfection of si-circ_0026416 or NC. g Schematic illustration of hsa_circ_0026416 pmirGLO luciferase reporter vectors. h Luciferase activity of WT or MUT pmirGLO-circ_0026416 after transfection with miR-346 mimics or NC in HEK293T cells. i Venn diagram for overlap among the four databases that predict miRNA target genes (miRDB, miRWalk, starBase v3.0, TargetScan 7.1). j Immunohistochemistry staining of NFIB in CRC tissues and normal colorectal mucosa from THPA data. k Difference in NFIB staining score in CRC tissues (n = 11) and normal tissues (n = 2) from THPA data. l qRT-PCR for expression of NFIB in 24 pairs of CRC and normal colorectal tissues. m qRT-PCR for expression of NFIB in HCT-8 or SW480 cells after transfection of si-circ_0026416 or NC. n Schematic illustration of NFIB 3′UTR pmirGLO luciferase reporter vectors. o Luciferase activity of WT or MUT pmirGLO-NFIB 3′UTR after transfection with miR-346 mimics or NC in HCT-8 and SW480 cells. p–s Western blot assays showed that overexpression of hsa_circ_0026416 promotes protein expression of NFIB, while knockdown of hsa_circ_0026416 inhibits NFIB expression in HCT-8 and SW480 cells. Data are presented as means ± SD of three independent experiments. ANM = adjacent normal mucosa, TCGA = The Cancer Genome Atlas, THPA = The Human Protein Atlas, NC = negative control. *P < 0.05, **P < 0.01, ***P < 0.001