Fig. 5

The validation of HK2 as the functional target gene for miR-654-3p. A Schematic plot showed the predicted miR-654-3p target sites in HK2 3’UTR WT and their mutations in HK2 3ʹUTR MUT. B, C Dual-luciferase reporter assay determined relative luciferase activity of HK2 3ʹUTR WT and HK2 3’UTR MUT report vectors in the condition of miR-654-3p or miR-NC overexpression in SKOV3 and A2780 cells. D QPCR estimated relative HK2 mRNA expression in tumor tissues and adjacent tissues from OVCA cancer patients (n = 52). E IHC showed the in situ expression of HK2 in one pair of adjacent and tumor tissues from one randomly selected OVCA patient. F Western blotting detected relative protein expression of HK2 in tissue extract of three pairs of adjacent and tumor tissues from three randomly selected OVCA patients. G–I Western blotting detected relative protein expression of HK2 in (G) HOSEpiC, SKOV3 and A2780 cells, and (H, I) SKOV3 and A2780 cells after transfections of si-NC, si-circPDXK#2, and si-circPDXK#2 combined with in-miR-NC or in-miR-654-3p. J, K Linear correlation coefficient (r) between HK2 mRNA level and miR-654-3p or circPDXK level in 52 OVCA tumors was determined by Pearson correlation analysis. **P < 0.01; ***P < 0.001