Fig. 3

DDIT4 was the direct target of miR-496 in PCa cells. a MiRanda found out seven hundred and fifteen mRNAs that might bind with miR-496. b RT-qPCR selected mRNAs whose expression conformed to following four conditions: highly expressed in PC3 cells, inhibited by miR-496 mimics, highly expressed in VCaP cells and suppressed by sh-NNT-AS1. c RT-qPCR examined the expression of DDIT4 in PCa cells. d ENCORI predicted the binding sites between DDIT4 and miR-496. e RNA pull down experiments demonstrated that the abundance of DDIT4 in indicated groups. f Luciferase reporter assays examined the relative luciferase activity in VCaP and PC3 cells with different transfections. g The expression of DDIT4 was detected via RT-qPCR in VCaP and PC3 cells transfected with shRNAs targeting DDIT4. H. CCK-8 assay indicated cell viability when DDIT4 was silenced in VCaP and PC3 cells. i, j EdU and colony formation assays detected the proliferation of VCaP and PC3 cells with DDIT4 inhibition. k, l Flow cytometry analysis and JC-1 assay evaluated the apoptosis of VCaP and PC3 cells with DDIT4 suppression. m Wound healing assay displayed the migration of VCaP and PC3 cells under DDIT4 down-regulation. *p < 0.05, **p < 0.01