Extended Data Figure 9. Antidotes work on further gut commensals, but do not compromise antibiotic efficacy on relevant pathogens.

a. 8 x 8 checkerboard assays to investigate if antidote is also protective for additional gut commensals. All combinations were tested in MGAM medium under anaerobic conditions. Heat map depicts bacterial growth based on median AUCs from 2-3 independent replicates. Concentrations are stated in μM.

b. 8 x 8 checkerboard assays to evaluate antidote effects on the activity of erythromycin and doxycycline in relevant pathogenic species. The gastrointestinal pathogens E. faecalis and E. faecium were tested under anaerobic conditions. S. aureus, a cause of extra-intestinal infections, such as bacteremia and infective endocarditis, was tested under aerobic conditions. Heatmaps depict mean normalized AUCs of three biological replicates. Antidotes exhibit either neutral or even slight synergistic effects with antibiotics.

c. Dicumarol rescues commensal growth (n=2, anaerobic conditions) in a concentration-dependent manner. Erythromycin still retains its activity against pertinent pathogens such as E. faecium, E. faecalis (n=3, anaerobic conditions) and S. aureus (n=3, aerobic conditions) - see Suppl. Table 1 for strains used. 0.65 μM (~0.5 μg/ml) erythromycin is within range of the MIC breakpoints for Staphylococcus (1 μg/ml) and Streptococci groups A, B, C & G (0.25 μg/ml). Error bars depict standard deviation.