Research Paper Volume 12, Issue 22 pp 22759—22775

Figure 7. NEAT1 up-regulated SMAD2 expression through sponging miR-200b-3p. (A) Spearman’s correlation analysis showed the correlation of NEAT1 and SMAD2 in malignant melanoma tissues (n=18) (R=0.5944, P < 0.001). (B–C) RT-qPCR and western blot were performed to assess mRNA and protein expression of SMAD2 after A375 cells were transfected si-NEAT1 plasmid or NC plasmid treated for 24 hours. (D) Luciferase reporter assay in HEK-293T cells transfected with luciferase report plasmids containing SMAD2 3’-UTR (pMIR-SMAD2 3’-UTR) and then were co-transfected with LV-miR-200b-3p or pCDNA3.1-NEAT1. miR-NC and pcDNA3.1-NC were used as control. (E–F) RT-qPCR and western blot were performed to assess mRNA and protein expression of SMAD2 after A375 cells were transfected with LV-NEAT1 and/or miR-200b-3p inhibitor or control.