Fig. 5

MiR-3173-5p binds to the 3'UTR of SOCS3 mRNA and inhibits its expression. A The binding sites of miR-3173-5p on SOCS3 mRNA 3'UTR region were predicted through Starbase database. Dual luciferase reporter assay was performed using pmirGLO-WT-SOCS3 and pmirGLO-MUT-SOCS3 reporter in the presence of miR-3173-5p mimic or miR-NC. Renilla luciferase control plasmids were co-transfected into cells, and the relative firefly luciferase activity in the reporter was normalized to renilla luciferase activity in the control plasmid. B SOCS3 protein levels were examined by Western blot after the transfection of miR-3173-5p mimic in PC-3 and LNCaP cells. C miR-3173-5p expression level was examined by RT-qPCR in PC-3 and LNCaP cells after the transfection with miR-3173-5p inhibitor or NC inhibitor. D SOCS3 protein levels were assessed by Western blot after the transfection with miR-3173-5p inhibitor or NC inhibitor. E Cells were transfected with pcDNA vector, pcDNA-LINC00893 plasmid or pcDNA-LINC00893 + miR-3173-5p mimic for 48 h, and SOCS3 protein levels were examined by Western blot. F SOCS3 protein levels in PCa cell lines and human normal prostate epithelial cell line RWPE-1 were evaluated by Western blot. G Relative SOCS3 mRNA levels in 66 pairs of PCa tissues and para-cancerous tissues were analyzed by RT-qPCR. H. SOCS3 protein levels in PCa tissues and matched para-cancerous tissues were evaluated by IHC. Scale bar: 100 μM. I, J The correlations between LINC00893 and SOCS3 mRNA, and between MiR-3173-5p and SOCS3 mRNA were analyzed by Spearman correlation coefficient analysis. Three independent assays were performed with three technical replicates. The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001