Fig. 4: CircUBE2J2 acted as a ceRNA to sponge miR-370-5p in HCC cells.

A RAP assays were performed using an biotin probe against circUBE2J2 on extracts from HepG2 cells. Relative expression levels of circRNA were evaluated by qRT-PCR. B The relative expression level of potential target miRNA of circUBE2J2 was assessed by qRT-PCR. C, D RIP experiments were performed using an antibody against AGO2 on extracts from HLF and HepG2 cells. E Schematic diagram of reporter gene structure. F A schematic drawing showing the putative binding sites. The mutant version of circUBE2J2 is presented. G, H Relative luciferase activity determined 48 h after transfecting HEK293T cells (G) and HepG2 cells (H) with miR-370-5P mimic/NC or circUBE2J2 WT/Mut. WT wild type, Mut, mutant-type. I The expression of circUBE2J2 in MHCC97H, HLF, and HepG2 cells after transfection with miR-370-5P mimcs or miR-370-5P inhibitor. J The expression of miR-370-5P in HepG2, HLF, and MHCC97H cells after transfection with circUBE2J2-OE or circUBE2J2 shRNA. K RAP assay were performed using an probe against miR370-5P on extracts from HepG2 cells. Relative expression levels of circRNA were evaluated by qRT-PCR. L FISH assay analysis for the co-localization of miR-370-5P and circUBE2J2 in HCC cells. M miR-370-5P co-localized with circUBE2J2 in HCC adjacent non-tumor and tumor tissues was detected by FISH. Data are from three independent experiments (mean ± SEM). The data are represented as the mean ± SD, n = 3. *p < 0.05; **p < 0.01; NS no significance.