Fig. 3: Divergent thymidine metabolism can be exploited using thymine supplementation to specifically target P. aeruginosa with fluorofolin.

a, Growth of E. coli MG1655 or b, P. aeruginosa PA14 in 0.3 mM TMI-supplemented media and treated with fluorofolin at 2× MIC or DMSO. Curves represent average OD600 of 2 biological replicates. c, Competition of P. aeruginosa PA14 and E. coli MG1655 in LB or TMI-supplemented LB media. PA14 and E. coli MG1655 were inoculated at a 1:1 ratio and grown overnight in the presence or absence of 50 μg ml−1 fluorofolin in each media condition. The following day, c.f.u.s were counted on LB agar or Pseudomonas selection agar plates to determine c.f.u. ml−1 of each species. Data represent mean ± s.d. of 3 biological and 3 technical replicates. ***P ≤ 0.001, P = 0.0006; calculated using two-sided unpaired t-test using Prism 9. d, Representative culture images.