Fig. 4: LOX-1 regulated intracellular H. pylori survival in gastric epithelial cells.

a, e Two individual siRNAs or overexpression plasmids were transfected into HFE145 cells to knock down or overexpress LOX-1. The knockdown or overexpression efficiency was validated by Western blots. GAPDH was served as a loading control. b–d, f–h HFE145 cells transfected with siRNAs or overexpression vector of LOX-1 were infected with H. pylori (MOI = 100) for 24 h. b, f Intracellular H. pylori 16S ribosome DNA levels were measured by real-time PCR. Human GAPDH was used as an internal control (n = 3 replicates for each group). c, g Cells were stained to visualize invaded H. pylori (Red) and nuclei (Blue). Ten visual fields of each group were randomly selected to count H. pylori number. d, h Cells were permeabilized with 1% saponin for 15 min. The diluted samples were then spread on H. pylori-selective blood agar plates and incubated for 5 days to count the colony number (n = 3 replicates for each group). i HFE145 cells transfected with WTAP siRNAs and/or LOX-1 siRNAs were infected with H. pylori (MOI = 100) for 24 h. Intracellular H. pylori 16S ribosomal DNA levels were measured by real-time PCR. Human GAPDH was used as an internal control (n = 3 replicates for each group). j, k Wild type (n = 20 animals) and Lox-1^−/− mice (n = 18 animals) were infected with H. pylori strain SS1 for 1 month. j Mouse stomach tissues were harvested and weighted, and then homogenized in sterile PBS. The samples were further diluted and spread on H. pylori-selective blood agar plates. After growing for 5 days, the colony numbers were count for quantitative analysis. k Parafilm-embedded sections of mouse stomach were stained to visualize Lox-1 (green), H. pylori (Red) and the nuclei (Blue). Ten visual fields of each sample were randomly selected to count H. pylori number and measure the colocalized signals of H. pylori and Lox-1. Scale bar, 10 μm. All the quantitative data were shown as means ± SD. Statistical analysis of the data was performed using unpaired two-tailed Student’s t test (f, g, h, j, k) or one-way ANOVA (b–d, 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 (b–d, f, g–i) was calculated from one of three independent experiments with similar results. Source data are provided as a Source Data file.