Extended Data Fig. 1: H. pylori infection increases ROS accumulation, decreases GSH levels, and enhances protein cysteine sulfenylation in AGS cells.
From: An infection-induced oxidation site regulates legumain processing and tumor growth

(a) Intracellular ROS in AGS cells incubated with H. pylori G27MA (MOI 50, 18 h), 5 mM hydrogen peroxide for 1 h, or media alone were quantified using the fluorogenic ROS indicator CM-H2DCFDA (left). Fluorescence measurements were normalized by the total number of live cells per condition (right). (b) Intracellular GSH levels in H. pylori-infected (H. pylori G27MA, MOI 50, 18 h) and uninfected AGS cells (left), normalized by the total number of live cells per condition (right). Data represent three independent experiments. Each circle represents an independent experiment. Bars represent means ± SD. *P = 0.03; **P = 0.007; ***P = 0.005; ****P < 0.0001; n.s., not significant. Two-way analysis of variance (ANOVA) with Å Ãdák’s multiple comparisons test was used for (a); two-tailed unpaired t-test was used for (b). (c) Western blot analysis of biotinylated proteins in H. pylori-infected (H. pylori G27MA, MOI 25, 18 h) and uninfected AGS cells. Proteins labeled with the sulfenic acid-specific probe DYn-2 were conjugated with diazo biotin azide and enriched on streptavidin beads prior to Western blot analysis. Western blot analysis was performed twice with consistent results.