FIG 6.

DMS3_acrIF1 ameliorates P. aeruginosa antibiotic resistance. (A) PA14 (OD₆₀₀ = 0.5) was evenly spread on a plate containing different antibiotics (Amp⁺: 100 μg/mL, Kan⁺: 100 μg/mL, Gen⁺: 100 μg/mL, Str⁺: 100 μg/mL), and antibiotic resistance was tested by observing the growth of bacteria on the plate. (B) PA14 (5 × 10⁸ CFU/mL) was cultured in lysogeny broth (LB) containing different antibiotics (Amp⁺: 100 μg/mL, Kan⁺: 100 μg/mL, Gen⁺: 100 μg/mL, Str⁺: 100 μg/mL) or treated with 1 × 10⁸ PFU/mL EATPs/DMS3 (MOI = 0.2). The growth kinetics of PA14 were continuously monitored at 4 h intervals up to 48 h at 37°C with constant shaking. (C) The left panel is a schematic diagram of the antibiotic resistance experiment. PA14 (PA14: 5 × 10⁸ CFU/mL) was treated with gradient concentrations of antibiotic (Gen⁺ or Str⁺), gradient concentrations of antibiotic and DMS3 (MOI = 0.02) (Gen⁺/Str⁺ + DMS3) or Gen⁺/Str⁺ + DMS3_acrIF1. The growth kinetics of PA14 were measured. (D) PA14 that acquired antibiotic resistance were picked out and grown to the mid-logarithmic phase (OD₆₀₀ = 0.4 to 0.6) in LB at 37°C with 220 rpm shaking. The growth kinetics of the acquired antibiotic resistance of PA14 were measured after treatment with DMS3_acrIF1 (MOI = 0.2) or with 100 μg/mL Gen^+/Str⁺ antibiotics. Data were presented as mean ± standard error of the mean (SEM), determined from biological triplicates and analyzed via a one-way ANOVA compared against a control group (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).