Fig. 1: Bioenergetic stress potentiates antimicrobial resistance evolution and persistence.
From: Bioenergetic stress potentiates antimicrobial resistance and persistence
a Energetic metabolite profiles for untreated (UT) and ciprofloxacin treated (CIP) E. coli MG1655 cells as determined by LC-MS/MS. Exponential phase cells were grown in MOPS minimal media and treated with ± 16 ng/mL ciprofloxacin for 1 h (n = 6). (p = 0.0066 for [ATP], 0.0335 for ATP/ADP, 0.0186 for adenylate energy charge [AEC], 0.0074 for [NADH], and 0.0807 for NADH/NAD+ by two-sided unpaired t-tests). b Energetic metabolite profiles for pEmpty, pF1, and pNOX cells grown in MOPS-rich media as determined by LC-MS/MS or Promega NAD/NADH-Glo (n = 4). Statistical testing by one-way ANOVA with Dunnett’s multiple comparisons test versus pEmpty for [ATP] (p = 0.0035; pF1 p = 0.0088, pNOX p = 0.0030), ATP/ADP (p = 0.0014; pF1 p = 0.0008, pNOX p = 0.0318), AEC (p = 0.0055; pF1 p = 0.0037, pNOX p = 0.0227), [NADH] (p < 0.0001; pF1 p = 0.0012, pNOX p < 0.0001), and NADH/NAD+ (p < 0.0001; pF1 p = 0.0007, pNOX p < 0.0001). c Growth curves for pEmpty, pF1, and pNOX cells grown in MOPS-rich media (n = 4). d Exponential phase oxygen consumption rates (OCR) as a reporter of respiratory activity (n = 7 for pEmpty and pF1; n = 8 for pNOX). Statistical testing by one-way ANOVA (p = 0.0011) with Dunnett’s multiple comparisons test versus pEmpty (pF1 p = 0.0008, pNOX p = 0.0068). e Exponential phase extracellular acidification rates (ECAR) as a reporter of glycolytic activity (n = 7 for pEmpty and pF1; n = 8 for pNOX). Statistical testing by one-way ANOVA (p = 0.0097) with Dunnett’s multiple comparisons test versus pEmpty (pF1 p = 0.0050, pNOX p = 0.1621). f Growth of OD600 = 0.0001 exponential phase cells in the presence of ciprofloxacin (n = 4). g Ciprofloxacin resistance evolution for pEmpty, pF1, and pNOX cells over 8 cycles of serial-passage (n = 12). Data reported as change in the minimum concentration for 50% growth inhibition (MIC50) relative to Cycle 1. Statistical testing by two-way ANOVA (p = 0.0150) with Dunnett’s multiple comparisons test versus pEmpty (Cycle 1: not determined; Cycle 2: pF1 p = 0.1097, pNOX p = 0.0490; Cycle 3: pF1 p = 0.0448, pNOX p = 0.3654; Cycle 4: pF1 p = 0.0170, pNOX p = 0.1550; Cycle 5: pF1 p = 0.0012, pNOX p = 0.1447; Cycle 6: pF1 p = 0.0014, pNOX p = 0.0817; Cycle 7: pF1 p = 0.0099, pNOX p = 0.2457; Cycle 8: pF1 p = 0.0429, pNOX p = 0.4146). h Ciprofloxacin lethality following treatment with 18 ng/mL ciprofloxacin (n = 4). Data reported as change in colony-forming units (CFUs) relative to time 0. Data depicted as mean ± 95% CI. Statistical testing by two-way ANOVA (p < 0.0001) with Dunnett’s multiple comparisons test versus pEmpty (0 h: pF1 p = 0.9986, pNOX p = 0.9722; 1 h: pF1 p = 0.7873, pNOX p = 0.1278; 2 h: pF1 p = 0.0007, pNOX p = 0.0021; 3 h: pF1 p < 0.0001, pNOX p = 0.0004; 4 h: pF1 p = 0.0002, pNOX p = 0.0014). All experiments performed in MOPS-rich media unless otherwise indicated. All data represent biological replicates (independent cultures) and are depicted as mean ± SEM unless otherwise indicated. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. p ≥ 0.1 not shown. Source data are provided within the ‘Source Data.xlsx’ file.
