Fig. 5

miR-194-5p targets NUDC in RCC. A Schematic of our strategy to identify the downstream target of miR-194-5p. B Pearson correlation analysis demonstrated that the expression of NUDC was negatively correlated with that of PUS10 in KIRC based on the TCGA database. (r = − 0.34, P value < 0.0001). C, D Changes in the expression of NUDC at the mRNA and protein levels upon transfection of miR-194-5p mimics and inhibitor in RCC cells were determined by qRT‒PCR and western blotting. Data from three independent experiments are presented as the mean ± SD in the histogram. E RIP assays were conducted to demonstrate the enrichment of NUDC on an anti-Ago2 antibody, suggesting that its degradation might depend on Ago2 and microRNAs. Data from three independent experiments are presented as the mean ± SD in the histogram. F Dual luciferase reporter assays were conducted by transfecting plasmids containing the WT or Mut 3ʹUTR of NUDC and miR-194-5p or control mimics into HEK-293T cells. The relative luciferase activity was normalized to Renilla luciferase activity. Data from three independent experiments are presented as the mean ± S.D. G, H qRT‒PCR was performed to measure the changed expression of NUDC mRNA upon the silencing or ectopic expression of PUS10 in Caki-1 and 786-O cells. Data from three replicated experiments are presented as the mean ± SD in the histogram. I Western blot results reflected the repressed expression of NUDC upon the ectopic expression of wild-type or mutant incompetent PUS10 in Caki-1 and 786-O cells. G Western blot assay showed that the increased expression of NUDC induced by PUS10 depletion could be reversed by miR-194-5p mimics. *P < 0.05, **P < 0.01, ***P < 0.001; ns not significant