Fig. 5

The expression of Semaphorin 4D (simplified as SEMA4D) in ESCC tissues and cells. a The miRDB predicted a complementary site between the wild type of SEMA4D 3′UTR (simplified as SEMA4D-wt) and miR-595. SEMA4D-wt was mutated of this potential binding sequence, named as SEMA4D-mut. b, c Dual-luciferase reporter assay confirmed luciferase activity of KYSE30 and KYSE450 cells co-transfected with SEMA4D-wt/mut and miR-595 or miR-NC. d–f RT-qPCR detected d miR-595 and e SEMA4D mRNA expression levels, and western blotting measured SEMA4D protein expression level in KYSE30 and KYSE450 cells introduced with miR-NC, miR-595, anti-miR-NC, or anti-miR-595. g RT-qPCR detected SEMA4D mRNA expression level in Tumor (n = 42) and Normal (n = 42) from ESCC patients. h Western blotting measured SEMA4D protein level in three paired tissues from ESCC patients. i Pearson’s correlation coefficient assay analyzed the correlation between miR-595 and SEMA4D mRNA expression in ESCC tumor tissues (n = 42). j, k RT-qPCR and western blotting detected SEMA4D expression levels in HET-1A, KYSE30 and KYSE450 cells. *P < 0.05