To achieve NELFE overexpression or knockdown in GC cell lines, NELFE-overexpression plasmid (pNELFE) was transfected into MGC-803 and HGC-27 cells with a relatively low NELFE expression, and specific siRNAs against NELFE (siNELFE-1 and siNELFE-2) were constructed and transfected into BGC-823 and AGS cells with high levels of NELFE expression, after which western blot analyses confirmed the transfection efficiencies (Fig. 2a). Here, NELFE is linked to gastric cancer.