Fig. 1. Tolerance evolves during repeated SDM but not WDM antibiotic treatment.

a Schematic of evolution. An overnight culture of E. coli BW25113 was diluted 1:100 in LB, then incubated with an SDM or WDM antibiotic at the indicated concentration for 6 hours in a deep 96-well plate in triplicate. Next, plates were centrifuged and washed two times in PBS. Cells were then re-suspended in fresh LB, grown overnight, and the process was repeated the following day. b Percent survival of ancestral and evolved cells against SDM and WDM antibiotics. Horizontal axis labels indicate the antibiotic on which the culture was evolved and the evolution day (i.e., amp D6 indicates cells were evolved on ampicillin for 6 days). Data are representative of three biological replicates; error bars show SEM. c MICs of ancestral and evolved cultures. Experiments were performed in biological triplicate; error bars indicate SEM. d Cross-tolerance of ampicillin- and ciprofloxacin-evolved cultures. Horizontal axis labels indicate the antibiotic tested for cross-tolerance. The figure legend indicates the antibiotic on which the culture was evolved and the evolution day. Shown is the mean of three biological replicates; error bars indicate SEM. Source data are provided as a Source Data file.