Fig. 6

Effects of cHP1BP3 on malignant behavior of BLCA cells. A Venn diagram of predicted circRNAs acting as miR-1-3p sponges, from miRanda, RNAhybrid, and TargetScan databases. B Venn diagram of identified upregulated circRNAs and predicted circRNAs acting as miR-1-3p sponges. C RT-qPCR analysis of circRNA levels in streptavidin-captured fractions from YTS-1 cell lysates following transfection with 3′-end biotinylated miR-1-3p or control RNA (NC). **, p < 0.01. D Confirmation by luciferase reporter assay of miR-1-3p-binding sites on circRNAs. Control miRNA or miR-1-3p mimics were co-transfected with luciferase reporters containing predicted miR-1-3p binding sites. *, p < 0.05; **, p < 0.01; ***, p < 0.001. E Luciferase activity of luciferase-cHP1BP3 or its mutant in HEK293 cells following co-transfection with miR-1-3p. ***, p < 0.001. F Stability of cHP1BP3 following 24 h transcription blocking by actinomycin D (ActD) treatment, by RT-qPCR analysis. G Expression of cHP1BP3 amplified with random hexamer or oligo (dT)18 primers, by RT-qPCR. **, p < 0.01. H cHP1BP3 expression in BLCA and adjacent normal tissues. I-K Proliferation (I), migratory ability (J), and colony formation (K) of cHP1BP3-silenced YTS-1 vs. control cells. *, p < 0.05; **, p < 0.01; ***, p < 0.001. L-N Tumor size (L), volume (M), and weight (N) at wk. 3 after s.c. injection of cHP1PB3-silenced YTS-1 cells (n = 7). **, p < 0.01; ***, p < 0.001. O, P Microphotographs of splenic (O) and liver metastatic tumors (P)