Fig.1

MiR-659-3p targets SRPK1 gene 3’UTR and negatively regulates SRPK1 expression in osteosarcoma cells. A MiR-659-3p was downregulated in human osteosarcoma compared with normal bone cells. The data was analyzed from the existing dataset from NCBI Gene Expression Omnibus GSE28423. B Quantitative real-time PCR verified that downregulated expression of miR-659-3p in human osteosarcoma cell lines compared with normal bone cells (hFOB1.19). C Quantitative real-time PCR showed that relative mRNA expression of SRPK1 is upregulated in osteosarcoma cell lines compared with normal bone cells. D Reverse correlation between miR-659-3p expression and SRPK1 mRNA expression in the cell lines assayed. E Worse prognosis observed in low miR-659-3p expression osteosarcoma patients compared with high miR-659-3p expression patients. F Alignment of miR-659-3p and SRPK1 3’ UTR showed potential targeting sites. G Western blot showed SRPK1 protein expression in human osteosarcoma cell lines, which confirmed the rough reverse correlation with miR-659-3p expression in the cell lines. SRPK1 and GAPDH western blots were performed on different gels. H Luciferase assay confirmed that miR-659-3p targets SRPK1 3’ UTR and negatively regulated its expression. ** indicates P < 0.01 and *** indicates P < 0.001 compared with control