Fig. 1

Subcellular localization of CAG and CPG in AGS cells. a Biosynthetic pathway for cholesterol α-glucoside derivatives in H. pylori. b Representative images of puncta in AGS cells. AGS cells were treated with CG-MAN, CAG-MAN, CPG-MAN, or MAN (green), fixed, and imaged with confocal microscopy. Scale bar: 5 μm. c, d CAG-MAN and CPG-MAN are internalized to early endosomes and subsequently routed to lysosomes. AGS cells were treated with CAG-MAN or CPG-MAN, followed by immunostaining for EEA1 (early endosome antigen 1, red) or Lysotracker Red™ (red) to detect early endosomes (c) and lysosomes (d), respectively. e Removal of membrane cholesterol abolishes CAG internalization. AGS cells were pretreated with (or without) methyl β-cyclodextrin (MβCD) to sequester membrane cholesterol for 1 h, stained with GM1 (red) for 30 min, washed three times, and treated with CAG-MAN (green) or CPG-MAN (green) for another 1 h. The cells were then fixed and imaged with confocal microscopy. Scale bar: 5 μm. f Overlapped images of H. pylori 26695 with the puncta of intracellular CAG-MAN or CPG-MAN. AGS cells were treated with CAG-MAN (green) or CPG-MAN (green) for 3 h and co-cultured with H. pylori 26695 for 2 h. The cells were then fixed and immunostained with anti-H. pylori (red). Arrowheads indicate co-localization of H. pylori with puncta. Scale bar: 5 μm