Fig 1: CircDYRK1A inhibits glutamine metabolism in GC cells. AGS cells were transduced with circDYRK1A overexpression vector and HGC-27 cells were transduced with sh-circDYRK1A. A The expression of glutamine in AGS and HGC-27 cells detected using the kit. B The expression of glutamic acid in AGS and HGC-27 cells detected using the kit. C The expression of a-KG in AGS and HGC-27 cells detected using the kit. D The protein levels of GLS and GDH in AGS and HGC-27 cells detected by Western blot analysis. The cell experiment was repeated three times. * p < 0.05 vs. sh-NC-treated HGC-27 cells, or Vector-treated AGS cells
Fig 2: CircDYRK1A inhibits the malignant phenotypes and glutamine metabolism in GC cells by upregulating the expression of FBXO4 through miR-889-3p. AGS cells were transduced with sh-FBXO4-1 or sh-FBXO4-2. A Expression of FBXO4 in AGS cells determined by RT-qPCR. AGS cells were transduced with sh-FBXO4 and/or circDYRK1A, and HGC-27 cells were transduced with oe-FBXO4 and/or sh-circDYRK1A. B Expression of FBXO4, circDYRK1A and miR-889-3p in AGS cells and HGC-27 cells determined by RT-qPCR. C Proliferation of AGS cells and HGC-27 cells detected by EdU assay. D Migration of AGS and HGC-27 cells detected by Transwell assay. E Invasion of AGS and HGC-27 cells detected by Transwell assay. F Expression of glutamine in AGS and HGC-27 cells measured using the kit. G Expression of glutamic acid in AGS and HGC-27 cells measured using the kit. H Expression of a-KG in AGS and HGC-27 cells measured using the kit. I Protein levels of GLS and GDH in AGS and HGC-27 cells measured by Western blot analysis. The cell experiment was repeated three times. * p < 0.05
Fig 3: miR-889-3p reverses the inhibitory effect of circDYRK1A on malignant phenotypes of GC cells. AGS cells were transduced with miR-889-3p mimic and/or circDYRK1A, and HGC-27 cells were transduced with miR-889-3p inhibitor and/or sh-circDYRK1A. A Expression of circDYRK1A and miR-889-3p in AGS and HGC-27 cells determined by RT-qPCR. B Proliferation of AGS and HGC-27 cells detected by EdU assay. C Migration of AGS and HGC-27 cells detected by Transwell assay. D Invasion of AGS and HGC-27 cells detected by Transwell assay. E Expression of glutamine in AGS and HGC-27 cells measured using the kit. F Expression of glutamic acid in AGS and HGC-27 cells measured using the kit. G Expression of a-KG in AGS and HGC-27 cells measured using the kit. H Protein levels of GLS and GDH in AGS and HGC-27 cells measured by Western blot analysis. The cell experiment was repeated three times. * p < 0.05
Fig 4: RUNX3 promotes the expression of circDYRK1A and inhibits the progression of GC. A The expression of RUNX3 in microarray dataset GSE49051. B The expression of RUNX3 in GC tissues and adjacent normal tissues detected by RT-qPCR (n = 50). C RUNX3 expression in GC tissues and adjacent normal tissues measured by immunohistochemistry. D Pearson correlation analysis of the correlation between RUNX3 expression and circDYRK1A expression in GC tissues. E The enrichment of RUNX3 in circDYRK1A promoter region detected by ChIP assay. F The effect of RUNX3 on circDYRK1A promoter activity. G Expression of RUNX3 and circDYRK1A in AGS and HGC-27 cells measured by dual luciferase reporter assay. H Tumor size (left panel), growth curve (middle panel) and weight (right panel) in each group of mice detected (n = 6). I The apoptosis of tumor cells detected by TUNEL staining. Cell nuclei were stained in blue by hematoxylin, and yellow–brown cells are positive cells. J The expression of GLS and GDH (n = 6) measured by immunohistochemistry. The cell experiment was repeated three times. * p < 0.05
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