Fig. 3

NISCH is the downstream mRNA of miR-3657. A DIANA database was utilized to screen out the potential target mRNAs, RRAGD, CDK16, ZNRF1 and NISCH. B QPCR was used to evaluate the expression of potential target mRNAs after the knockdown of TFAP2A-AS1 in AGS and NUGC4 cells. C Western blot analysis was used to assess the protein level of NISCH in AGS and NUGC4 cells after the knockdown of TFAP2A-AS1. D, E RNA-pulldown and dual luciferase reporter assays were used to verify the interaction between NISCH and miR-3657 in AGS and NUGC4 cells. F, G QPCR and western blot analyses was used to detect the level of NISCH in AGS and NUGC4 cells after the transfection of sh-NC, sh-TFAP2A-AS1-1, sh-TFAP2A-AS1-1 + inhibitor-NC or sh-TFAP2A-AS1-1 + miR-3657 inhibitor. The statistical analysis for B, D, F was one-way ANOVA, and for E was two-way ANOVA. β-actin was used as the internal reference for gene expression analysis. **P < 0.01