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Fig. 1 | Biomarker Research

Fig. 1

From: Loss of miR-637 promotes cancer cell stemness via WASH/IL-8 pathway and serves as a novel prognostic marker in esophageal squamous cell carcinoma

Fig. 1

WASH is a direct target of miR-637 in ESCC cells. A The database miRanda and TargetScan were used to predict candidate miRNAs targeting WASH. B Relative expression of WASH was analyzed by qRT-PCR in KYSE70 cells transfected with the miRNA mimics for negative control (NC) or predicted candidates targeting WASH. C, D The down-regulation of WASH expression was determined in both KYSE70 and KYSE450 cells transfected with miR-637 mimic by qRT-PCR (C) and Western blotting (D), respectively. E, F The up-regulation of WASH expression was confirmed in both KYSE70 and KYSE450 cells transfected with miR-637 inhibitor by qRT-PCR (E) and Western blotting (F), respectively. G, H The relative levels of miR-637 were measured by qRT-PCR assays in KYSE70 cells and KYSE450 cells after transfection with mi-637 mimic (G) or miR-637 inhibitor (H), respectively. I Schematic representation of WASH 3′-UTR wild type (WT), and mutant (Mut) with altered residues in the core region of miR-637 binding site. J Luciferase activity in 293 T cells co-transfected with WASH WT-3′-UTR or Mut-3′-UTR constructs and miR-637 or NC mimic, respectively. Data are presented as mean ± standard deviation. ns, not significant; *p < 0.05, **p < 0.01, ***p < 0.001

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