Fig. 4

ZFP64 mediated stemness enhancement in GC cells. A Heatmap showing changes in gene expression after the overexpression of ZFP64 in HGC-27 cells. B The enrichment of KEGG pathways and Gene Ontology (GO) terms among differentially expressed genes was determined by the DAVID Functional Annotation tool. C The GSEA revealed a significant enrichment of the upregulated stem cell signature gene set and downregulated apoptosis signature gene set in ZFP64-overexpressing HGC-27 cells. D-E qRT-PCR and western blot showing the levels of stemness-related genes in GC cells with different ZFP64 expression levels. F-G Levels of the stemness-related protein CD44 in GC cells were analyzed using flow cytometry. Blue represents the isotype control. H Sphere forming assay of GC cells with ZFP64 knockdown or overexpression. I ZFP64 overexpression enhanced tumor-initiating capacity in vivo, as analyzed by a limiting dilution assay. Data are presented as the means ± SEM. ** P < 0.01; *** P < 0.005