Fig. 2

Treatment of AGS cells with CAG increases the number of intracellular H. pylori. a Bacterial internalization is reduced in the absence of cholesterol glucosides. AGS cells were co-cultured with H. pylori 26695 or ΔCGT for 2, 4 or 6 h. The samples were then lysed at the indicated time points and plated on blood agar plates for counting viable colonies. Bacterial internalization is presented as the proportion of intracellular bacteria out of the total bacteria. b To study the effect on the number of intracellular bacteria, AGS cells were co-cultured with H. pylori 26695 or ΔCGT for 2, 4 or 6 h and then treated with gentamycin for 1 h. Samples were lysed, and lysates were plated on blood agar plates for colony counting to give the number of intracellular bacteria (CFU/ml). c Bacterial internalization depends on CAG. AGS cells were first treated with cholesterol, CG, CAG, or CPG for 1 h, and then infected with H. pylori 26695 or ΔCGT for an additional 3 h. Bacterial internalization was then measured as mentioned in (a). d To measure the number of intracellular bacteria, AGS cells were first treated with CG, CAG, or CPG and co-cultured with H. pylori 26695 or ΔCGT for 6 h, followed by gentamycin treatment. The number of intracellular bacteria was then measured as mentioned in (b). e GES-1 cells were treated with DMSO or CAG, and then infected with H. pylori 26695 or ΔCGT. The degree of bacterial internalization was measured as described in (a). f GES-1 cells were treated with DMSO or CAG, infected with H. pylori 26695 or ΔCGT, and then subjected to the measurement for the number of intracellular bacteria as described in (b). g Bacterial internalization was dependent on lipid rafts. After AGS cells were treated with MβCD and infected with H. pylori 26695 or ΔCGT for another 3 h, internalized bacteria were counted. The data shown in panels (a–c, e and g) are normalized relative values. h ΔCGT greatly reduced H. pylori colonization in the mice stomach. Six-week-old C57BL/6J mice were intragastrically infected with H. pylori 26695 or ΔCGT. At 10 weeks post-the infection, the stomach samples were processed for CFU enumeration as described in “Materials and methods”. The number of viable colonies were counted and shown as CFU/mg of mice stomach (n = 3). Data shown represent the mean ± SEM (standard error of the mean). **p < 0.01, *p < 0.05 vs. the control (n = 3); n.s. (not significant). i The cryosections of H. pylori 26695- and ΔCGT-infected mice stomach samples were stained with anti-H. pylori antibody (green) and for nucleus (blue), and imaged by confocal microscopy. Scale bar: 20 μm