Fig. 2: Validation of METTL3-CXCL1 Target Relationship.

A Western blot analysis of CXCL1 and METTL3 protein levels in GES-1 cells after co-culture with HP at different MOIs (0, 20, 50, 100, and 200) and for different durations (0, 3, 6, 12, and 24 h). B RT-qPCR analysis of CXCL1 and METTL3 mRNA levels in GES-1 cells after co-culture with HP at different MOIs (0, 20, 50, 100, and 200) and for different durations (0, 3, 6, 12, and 24 h). C RT-qPCR and Western blot screening of three si-METTL3 sequences. D MeRIP-qPCR experiment detecting m6A methylation modifications of CXCL1 mRNA in GES-1 cells. E RIP-qPCR experiment detecting the target relationship between CXCL1 mRNA and METTL3 in GES-1 cells. F Dual-luciferase reporter assay detecting the target relationship between CXCL1 and METTL3 in GES-1 cells. G Western blot analysis of CXCL1 and METTL3 protein expression levels in HP-infected GES-1 cells. H RT-qPCR analysis of CXCL1 and METTL3 mRNA levels in HP-infected GES-1 cells. I Immunofluorescence staining detecting the expression levels of CXCL1 and METTL3 in HP-infected GES-1 cells. J MeRIP-qPCR experiment detecting m6A methylation modifications of CXCL1 mRNA in GES-1 cells. K RIP-qPCR experiment detecting the target relationship between CXCL1 mRNA and BHLHE41 in GES-1 cells. L Dual-luciferase reporter assay detecting the target relationship between CXCL1 and BHLHE41 in GES-1 cells. M Western blot analysis of CXCL1 and BHLHE41 protein expression levels in HP-infected GES-1 cells. N RT-qPCR analysis of CXCL1 and BHLHE41 mRNA levels in HP-infected GES-1 cells. O Immunofluorescence staining detecting the expression levels of CXCL1 and BHLHE41 in HP-infected GES-1 cells. Scale bar: 25 μm. Values are presented as the mean ± standard deviation, and all cell experiments were repeated three times. ***indicates P < 0.001.