Fig 1: The effect of circ_1306/miR-584-5p/CDK16 on cell apoptosis in HCC cell lines. (A) Cell apoptosis was analysed by flow cytometry assay after transfection. **P < 0.01 compared to si-NC group. (B) The protein expression of apoptosis-related genes was treated by Western blot. **P < 0.01 compared to control group. ##P < 0.01 compared to si-1302 gourp. ??P < 0.01 compared to OV-CDK16 group. All data represent the mean ± SD
Fig 2: CDK16 was a direct target of miR-584-5p. (A) A Venn drawing showed the overlapping of the target genes of miR-584-5p predicted by GSE101728, miRDB, TargetScan and starbase. (B) The expression of CDK16 in HCC tissues (n = 7) and adjacent normal tissues (n = 7) in GSE108724. **P < 0.01 compared to adjacent normal tissues. (C) The binding site between miR-584-5p and CDK16 3’-UTR. (D) Dual-luciferase reporter assay showed that miR mimics significantly reduced luciferase activity of WT-CDK16 group. **P < 0.01 compared to miR-NC group. (E) The protein expression of CDK16 in the human normal liver cell line (L02) and HCC cell lines (SK-HEP-1, Hep G2, Huh-7 and Hep 3B) were determined by Western blot. **P < 0.01 compared to L02. (F) The protein expression of CDK16 in Huh-7 and Hep 3B cells after si-1306, miR-inhibitor, OV-CDK16 or co-transfection was tested by qPCR. **P < 0.01 compared to control group. ##P < 0.01 compared to si-1302 group. ??P < 0.01 compared to OV-CDK16 group. All data represent the mean ± SD
Fig 3: The effect of circ_1306/miR-584-5p/CDK16 on cell proliferation in HCC cell lines. (A) Cell viability was tested by CCK-8 assay after transfection. (B) Cell proliferation was tested by colony formation assay. **P < 0.01 compared to si-NC group
Fig 4: The effect of circ_1306/miR-584-5p/CDK16 on tumour growth in vivo. (A-C) si-1306 and miR mimics reduced xenograft growth in nude mice. (D-F) QRT-PCR and Western blot were used to explore circ_1306, miR-584-5p and CDK16 expression in tumour tissues. All data represent the mean ± SD. **P < 0.01 compared to si-NC group. (G) The expression of proliferation marked (Ki67) in tumour tissues was tested by IHC. (H) The cell apoptosis in tumour tissues was tested by Tunel assay
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