Extended Data Fig. 5. Time-course accumulation of Debio-1452 and derivatives in E. coli MG1655.

Bacterial cells were washed with and suspended in PBS. Cells were exposed to compound (50 μM final) and incubated at 37°C with shaking for 10 min, 30 min, 1 h, and 2 h. All cells were viable at these time points. Extra-cellular compound was removed, cells were lysed, and amount of compound in lysate was quantified by LC-MS/MS. Accumulation reported in nmol per 10¹² colony-forming units (CFUs). Data shown represents the average of three independent experiments. Error bars represent the standard deviation of the mean. Measurements were compared by ordinary two-way ANOVA. Tukey’s multiple comparisons test was used to compare compounds at each timepoint. Statistical significance from Debio-1452 is indicated with asterisks (ns, not significant when p > 0.05 (Debio-1452-NH3, 10 min, p = 0.1719; Debio-1452-NH3, 0.5 h, p = 0.5522), *** p < 0.001, **** p < 0.0001).