Fig. 4

miR-363 downregulates S1PR1 expression by directly targeting its 3′-UTR. a S1PR1 mRNA level changes in 786O and 769P cells treated with different interferences. b, c Overexpression of miR-363 decreased the expression of S1PR1 protein levels in ccRCC cells, whereas knockdown of miR-363 increased the expression of S1PR1 protein levels in ccRCC cells. d Representative immunofluorescent staining images showed the inverse effect of miR-363 on S1PR1 in ccRCC cells. e Sequence alignment of the S1PR1 3′-UTR with wild-type (WT) versus mutant (MUT) predicted potential miR-363 binding sites. f Luciferase reporter assay showed attenuated reporter activity after transfection of the wild-type S1PR1 3′-UTR reporter construct in human embryonic kidney 293T cells overexpressing miR-363. The S1PR1 3′-UTR MUT and control constructs had no effect on reporter activity. Data are presented as the mean ± SD. (*P < 0.05, **P < 0.01, ***P < 0.001)