All bacterial strains used in this study are listed in Supplementary file 1 . Escherichia coli strains were grown in Luria–Bertani broth (LB; Difco, France) or Luria–Bertani agar (LA; Difco) medium at 37°C with aeration. When needed, the antibiotics ampicillin (Amp; Sigma-Aldrich, Germany) and erythromycin (Ery; Sigma-Aldrich) were added at a final concentration of 100 µg/ml, and kanamycin (Km; Sigma-Aldrich) was added at a final concentration of 30 µg/ml. Lactococcus lactis LL108 strain was grown in M17 broth (Difco), supplemented with sucrose (0.5 M) and glucose (0.5% wt/vol) at 30°C without aeration. Erythromycin was used when required at 100 µg/ml. S. aureus strains were grown at 30°C with aeration in tryptic soy broth (TSB; Difco) or on tryptic soy agar (TSA; Difco). Medium was supplemented when required with Ery at 10 µg/ml and/or 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-Gal; Apollo Scientific, UK) at 100 µg/ml. S. pneumoniae was grown in C + Y medium at 37°C, without aeration, or on TSA plates supplemented with 5% vol/vol sheep blood (Probiológica, Portugal). Tetracycline (Sigma-Aldrich) and Ery were used, when required, at 1 µg/ml and 0.25 µg/ml, respectively.
Erythromycin ery
Manufactured by Merck Group
Sourced in Germany, Japan, United States
Erythromycin (ERY) is a macrolide antibiotic used in laboratory settings. It is effective against a range of gram-positive bacteria and some gram-negative bacteria. Erythromycin functions by inhibiting bacterial protein synthesis, thereby preventing the growth and reproduction of bacterial cells.
Automatically generated - may contain errors
Lab products found in correlation
Ampicillin amp, by Merck Group (2 mentions)
Tetracycline, by Merck Group (1 mentions)
Kanamycin km, by Merck Group (1 mentions)
Chloramphenicol cm, by Merck Group (1 mentions)
Streptomycin str, by Merck Group (1 mentions)
Trolox c, by Merck Group (1 mentions)
Cycloheximide chx, by Bio-Techne (1 mentions)
Tetracycline tet, by Merck Group (1 mentions)
Kanamycin kan, by Merck Group (1 mentions)
Ciprofloxacin cip, by Merck Group (1 mentions)
Gentamicin gen, by Merck Group (1 mentions)
Oxacillin oxa, by Merck Group (1 mentions)
Vancomycin van, by Merck Group (1 mentions)
6 protocols using erythromycin ery
1
Bacterial Strain Cultivation Protocols
2
Listeria monocytogenes and Listeria ivanovii Strains
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All the strains and plasmids used in this study are listed in Table S1 . Listeria monocytogenes 10403S and Listeria ivanovii PAM55 were provided by Dr. Hao Shen (Department of Microbiology, Perelman School of Medicine, University of Pennsylvania). Listeria strains were routinely cultured in brain/heart infusion (BHI) medium (Beijing LuQiao Company) at 37°C. All restriction enzymes were purchased from (NEB England). Erythromycin (Ery; Sigma‐Aldrich) was added at a concentration of 3 μg/ml. D‐alanine (J&K Scientific) was added to the BHI medium at a final concentration of 2 mg/ml (referred to as the D‐BHI plate or broth).
Female C57BL/6 mice, aged 6–8 weeks, were purchased from the Beijing Charles River Animal Laboratory. All mice were maintained under specific‐pathogen‐free (SPF) conditions throughout the experiments at the Animal Center of the School of Public Health at Sichuan University, and the experimental protocols were approved by the Animal Care and Use Committee of Sichuan University.
Female C57BL/6 mice, aged 6–8 weeks, were purchased from the Beijing Charles River Animal Laboratory. All mice were maintained under specific‐pathogen‐free (SPF) conditions throughout the experiments at the Animal Center of the School of Public Health at Sichuan University, and the experimental protocols were approved by the Animal Care and Use Committee of Sichuan University.
3
Bacterial Strains and Plasmids Protocol
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The bacterial strains and plasmids used in this work are listed in Table 1 . Escherichia coli TG1 and E. coli TOP 10 were grown aerobically in Luria-Bertani (Acumedia) medium (LB) at 37°C with shaking. L. lactis FnBPA+(Que et al., 2001 (link)) was grown in M17 (Difco, Sparks, MD, United States) medium supplemented with 0.5% glucose (GM17) at 30°C without shaking. When required, recombinant bacteria were selected by addition of antibiotics: for L. lactis FnBPA+(pValac:Ag85A) erythromycin (Ery, Sigma–Aldrich) at 5 μg/mL and chloramphenicol (Cm, Sigma–Aldrich) at 5 μg/mL; for E. coli (pValac:Ag85A), Cm at 10 μg/mL were used.
4
Preparation and Handling of Experimental Compounds
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The TC antibiotics, DOX , DMC, and CT , the inhibitor of lipid peroxidation Trolox-C (TROL; #238813), the iron chelator desferrioxamine (DES; D9533), the hydrogen peroxide detoxifying enzyme catalase from bovine liver (CAT; C100), vitamin C (VitC; 255564), and the two non-TC antibiotics erythromycin (ERY; E5389) and streptomycin (STR; S6501) were all from Sigma-Aldrich. The inhibitor of eukaryotic translation, the antifungal antibiotic cycloheximide (CHX; #0970), and the inhibitor of ferroptosis Liproxstatin-1 (LIP; #6113) were from Tocris Biotechne (Abingdon, UK). The two TCs, DDMC and DDOX, were obtained through in-house synthesis.
Stock solutions ofDOX and CT were made at 10 mM in distilled water and DMSO, respectively. Stock solutions of other TCs, DDOX , DMC, and DDMC, were prepared at 50 mM in DMSO. All stock solutions were stored at −20 °C. Intermediate dilutions of TCs used for cell culture treatments were made in distilled water and stored for 7 days at 4 °C, protected from light. Stock solutions of TROL and LIP were made at 50 mM in pure ethanol and DMSO, respectively. DES and VitC stock solutions were made at 10 mM in distilled water. CAT was prediluted at 5000 UI in distilled water and kept at 4 °C until use. Treatments with test compounds were carried out or renewed at the time of culture medium change.
Stock solutions of
5
Antimicrobial Susceptibility Assay
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CHL (Sigma-Aldrich, St. Louis, MI, USA) and thiamphenicol (TAP) (Wako Pure Chemical, Osaka, Japan) were dissolved in ethanol at a concentration of 20 mg/mL, whereas erythromycin (ERY) (Sigma-Aldrich) was dissolved in ethanol at a concentration of 10 mg/mL. Tetracycline (TET) (Sigma-Aldrich) was dissolved in ethanol at a concentration of 5 mg/mL. FLC, ITC, VRC, and MCZ were dissolved in dimethyl sulfoxide (DMSO) to constitute stock solutions of 0.5 mg/mL. Stock solutions were stored at −20 °C until use. Non-impregnated cellulose discs (BioMérieux, Marcy-l’Étoile, France) were used for the diffusion assays.
6
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility was tested by the agar dilution method following Clinical and Laboratory Standards Institute (CLSI) recommendations [29 ] for the following antibiotics: ampicillin (AMP; Sigma-Aldrich, St. Louis, MO, USA), oxacillin (OXA; Sigma-Aldrich), kanamycin (KAN; Sigma-Aldrich), gentamicin (GEN; Sigma-Aldrich), erythromycin (ERY; Sigma-Aldrich), clindamycin (CLI; Sigma-Aldrich), vancomycin (VAN; Sigma-Aldrich), ciprofloxacin (CIP; Sigma-Aldrich), and tetracycline (TET; Wako Pure Chemical Industries, Osaka, Japan). S. aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 served as quality control strains. The breakpoints of these antimicrobial agents were determined according to CLSI interpretation criteria [29 ].
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