a Diagram showing the identified Ag+-binding proteins in glycolytic pathway and how Ag+ disrupts glycolysis in S. aureus. G6P, glucose 6-phosphatase; F6P, fructose 6-phosphate; FBP, fructose 1,6-bisphosphate; GAP, glyceraldehyde 3-phosphate; 2PG, 2-phosphoglycerate; PEP, phosphoenolpyruvate; PYR, pyruvate; LAC, lactate. b Relative activities of glycolytic enzymes in S. aureus after treatment with 20 μg/mL AgNO3 for different time points (n = 3). c Relative ATP abundance of S. aureus after treatment with 0, 2, 10, 20, and 25 µg/mL AgNO3 (n = 3). The ATP contents in untreated S. aureus at corresponding time points were set as 100%. d Diagram showing how Ag+ disrupts electron transport chain and oxidative stress defense system in S. aureus. e Measurement of ROS levels with flow cytometry (n = 3). Left: CM-H2DCFDA fluorescence histogram of S. aureus with (yellow) or without (green) treatment of Ag+ (n = 3). Right: mean fluorescence intensity. f Diagram showing how Ag+ disrupts pentose phosphate pathway of S. aureus. G6P, glucose-6-phosphate; 6PGL, 6-phosphoglucono-δ-lactone; 6PG, 6-phosphogluconate; Ru5P, ribulose 5-phosphate; R5P, ribose 5-phosphate. g Expression level of Pgl and 6PGDH examined by western blotting (n = 3). Bar chart shows quantification of protein levels normalized to Gapdh and untreated control group. h Growth of WT S. aureus and mutants of pgl and gnd gene knock-down strains with or without treatment of 20 μg/mL AgNO3 (n = 3). i Relative activities of Pgl and 6PGDH in S. aureus after treatment with 20 μg/mL AgNO3 for different time points (n = 3). j NADPH/NADP+ ratio in S. aureus exposed to 20 μg/mL AgNO3 for various time points (n = 3). Mean value of three replicates is shown and error bars indicate ±SEM (b, c, e, g, h, i, j). Two-tailed t-test was used for all comparisons between two groups.