Fig. 5

The transcription factor ZFP64 regulates the transcription of GAL-1 in GC cells. A Heatmaps displaying ZFP64-binding sites in a 3 kb window around chromatin immunoprecipitation site followed by ChIP-seq peak summits in HGC-27 cells. B Overlap between RNA-seq and ChIP-seq data. C Representative browser track images showing the intensity of the ChIP-seq signal for GAL-1 in HGC-27 cells compared with the input control. D-E GAL-1 mRNA and protein were detected in GC cells with different ZFP64 expression levels. F The culture supernatants of GC cells were collected and analyzed for GAL-1 levels by ELIASA assay. G Pearson’s correlation analysis of the expression of ZFP64 and GAL-1 in GC tissues. H Serum levels of the GAL-1 protein in patients with GC. I Western plot analysis of ZFP64 and GAL-1 in GC tissues. J Relative luciferase reporter assays in HEK293 cells co-transfected with plasmid constructs containing the GAL-1 promoter with a ZFP64-overexpressing construct or control construct. K Relative luciferase reporter assays in the AGS, HGC-27, and MGC-803 cell lines after cotransfection of plasmid constructs containing the GAL-1 promoter with a ZFP64-overexpressing construct. L ZFP64-binding motif (upper panel); Serially truncated GAL-1 promoter constructs were transfected into HGC-27 cells. Then, pCMV-ZFP64 plasmids were co-transfected, and a luciferase reporter assay was conducted (lower panel). M ZFP64 active ERK1/2 and AKT signals, but have no effect on phosphorylation of p65. Data are presented as the means ± SEM. ** P < 0.01; *** P < 0.005