Fig. 3: LOX-1 was identified as a m⁶A-modified gene upon H. pylori infection.

a Integrated analysis of m⁶A-seq and RNA-seq from H. pylori-infected HFE145 cells transfected with negative control or WTAP siRNAs was shown. The fold change of m⁶A-seq was normalized to the corresponding transcript expression level. Genes with Log₂ fold change > 1 were highlighted. b The m⁶A peaks located at 3′UTR of LOX-1 in m⁶A-seq and the corresponding peaks in RNA-seq were visualized by IGV tool. c–e HFE145 cells transfected with negative control or WTAP siRNAs were infected with or without H. pylori for 24 h. c MeRIP-quantitative PCR was performed to validate m⁶A enrichment on LOX-1-3′UTR. The m⁶A enrichment of each group was calculated by m⁶A-IP/input (n = 3 replicates for each group). d LOX-1 mRNA levels of each group were measured by real-time PCR. Human ACTB was used as an internal control (n = 3 replicates for each group). e LOX-1 protein levels of each group were examined by Western blots. Human GAPDH was used as a loading control. f–h HFE145 cells transfected with negative control or siRNAs targeting METTL3/METTL14/WTAP were infected with H. pylori for 24 h. f LOX-1 mRNA levels of each group were measured by real-time PCR. Human ACTB was used as an internal control (n = 3 replicates for each group). g LOX-1 protein levels of each group were examined by Western blots. Human GAPDH was used as a loading control. h RNA decay rates of LOX-1 of each group were measured after treating with Actinomycin D (normalize to 0 h, n = 3 replicates for each group). i Two luciferase plasmids were constructed by inserting corresponding CDS into pmiR-RB-Report^TM vector. The wild type plasmid contained the full-length 3′UTR of LOX-1 and partial CDS near the stop codon, whereas five m⁶A-consensus motifs identified from m⁶A-seq were mutated with A-to-C conversion in the mutated plasmid. The relative luciferase activity was measured and calculated by normalizing Renilla to Firefly activity (n = 3 replicates for each group). All the quantitative data were shown as means ± SD. Statistical analysis of the data was performed using one-way ANOVA (c, d, f, h, i) followed by Tukey’s multiple comparison tests with adjustments and the corresponding p-values are included in the figure panels. The statistical significance of the data (c, d, f, h, i) was calculated from one of three independent experiments with similar results. Source data are provided as a Source Data file.