Supplementary Materials

Supplementary Material for:

Fitness cost of antibiotic susceptibility during bacterial infection

Damien Roux, Olga Danilchanka, Thomas Guillard, Vincent Cattoir, Hugues Aschard, Yang Fu, Francois Angoulvant, Jonathan Messika, Jean-Damien Ricard, John J. Mekalanos, Stephen Lory, Gerald B. Pier,* David Skurnik*

*Corresponding author. E-mail: gpier{at}bwh.harvard.edu (G.B.P.); dskurnik{at}rics.bwh.harvard.edu (D.S.)

Published 22 July 2015, Sci. Transl. Med. 7, 297ra114 (2015)
DOI: 10.1126/scitranslmed.aab1621

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Evolution of the RPKM sequencing reads for Tn insertions in genes in each of 27 functional classes from LB to the lung after 1 hour (A), from 1 to 6 hours in the lung (B), and from 6 to 24 hours in the lung (C).
  • Fig. S2. In vivo fitness of P. aeruginosa mutants with Tn insertions in genes from the functional class “motility and attachment” after 1, 6, or 24 hours of infection in the lung.
  • Fig. S3. Comparative in vivo fitness (in the GI tract and lung) of bacterial strains with Tn insertions in genes needed to produce T4aP components.
  • Fig. S4. Comparative in vitro fitness in LB and water of bacterial strains with Tn insertion mutants in genes needed to produce T4aP components.
  • Fig. S5. Comparative in vivo fitness of bacterial strains with Tn insertions in genes that encode all of the annotated VFs of P. aeruginosa (after 1, 6, or 24 hours in the lung).
  • Fig. S6. Evolution over time of the RPKM for Tn-interrupted genes in the LPS O-antigen locus.
  • Fig. S7. Increased virulence of oprD mutant carbapenem-resistant clinical strains of P. aeruginosa (48.2 and 51.2) in lung infection compared to isogenic (48.1 and 51.1) and complemented strains (48.2::PoprD and 51.2::PoprD).
  • Fig. S8. Evolution over time of the changes in the RPKM for Tn-interrupted genes associated with constitutive antibiotic resistance in P. aeruginosa in the GI tract, spleen, and lung.
  • Fig. S9. Analysis of in vitro growth and survival of Tn mutants deficient in genes associated with constitutive antibiotic resistance in P. aeruginosa.
  • Fig. S10. Effect of deletion of intrinsic antibiotic resistance genes on survival of P. aeruginosa in J774 macrophages.
  • Table S1. Analysis of selective pressures detected by RPKM reads during P. aeruginosa lung infection.
  • Table S2. Genes (116) whose loss shows an increased fitness for lung infection based on having Tn insertions with at least 100 reads after 24 hours and with reads further increasing more than twofold between 24 and 48 hours of infection.
  • Table S3. Antibiotic sensitivity results for wild-type P. aeruginosa PA14 and strains deleted for genes encoding intrinsic antibiotic resistance.
  • Table S4. Antibiotic sensitivity results for wild-type P. aeruginosa PA14 and strains deleted for genes encoding intrinsic antibiotic resistance (inhibition diameter).
  • Table S5. Antibiotic sensitivity results for wild-type P. aeruginosa PA14 and strains deleted for genes encoding intrinsic antibiotic resistance (MIC as measured by E test).
  • Table S6. Primers for genomic amplification used in this study.
  • Table S7. Bacterial strains used in this study.
  • Table S8. Plasmids used in this study.
  • Data file S1. Analysis of the TnSeq data by functional classes of P. aeruginosa PA14.
  • Data file S2. Fitness of flagellin mutants identified in the study.
  • References (4959)

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