Fig. 4

SMAD4 is the target gene of miR-452-5p and associated with good prognosis of RCC. a The schematic illustration of the proposed model depicting of the putative downstream target gene, SMAD4, of miR-452-5p from 4 prediction databases, miRTarBase, miRTar, TargetMiner and miRwalk. b SMAD4 expression in RCC and normal renal tissues from TCGA RCC dataset. c IHC staining revealed the level of SMAD4 expression in RCC tissues (×200, × 400) versus adjacent normal tissues. d SMAD4 expression and paitients’ survival time from TCGA RCC dataset according to www.proteinatlas.org. e the correlation between miR-452-5p expression and corresponding SMAD4 expression in RCC and normal renal tissues from TCGA RCC dataset. f The schematic illustration of predicted binding sites on the 3’-UTRs of SMAD4 mRNA and sequences of the wild-type (SMAD4 WtA, WtB, WtC) and mutated 3’-UTR-Renilla luciferase reporters (SMAD4 mutA, mutB, mutC). g Luciferase reporter assays of mutAB, BC, AC and ABC in OSRC-2 cells. h-j Western blot analysis for SMAD4 and SMAD7 protein levels of miR-452-5p comparing to miR-NC (h), anti-miR-452-5p comparing to anti-miR-NC (i) and LNA-miR-452-5p comparing to LNA-miR-NC (J) transfection in OSRC-2 and SW839 cell lines. k IHC staining revealed the level of SMAD7 expression in RCC tissues (×200, × 400) versus adjacent normal tissues. Data shown are mean±S.D. *p<0.05