Role of the serine protease Sat in the pathogenesis of sepsis caused by Escherichia coli

EC071 25% serum resistance – CFU.mL-1 counting (.pdf): colony forming units counting for bacterial survival after serum resistance assay. EC071 25% serum resistance – graph (.pdf): graphic representation of colony forming units counting for bacterial survival after serum resistance assay. EC071 plasmid analysis (.pdf): plasmid extraction analysis in an agarose gel. Genetic constructions and modifications – agarose gels: PCR, digestion and plasmid extraction products analysis in agarose gels. Growth curve – CFT073 (.pdf): growth curve of CFT073 in LB medium. Growth curve – EC071 and derivative strains (.pdf): growth curve of EC071 and derivative strains in LB medium. Growth curve – MG1655 and derivative strains (.pdf): growth curve of MG1655 and derivative strains in LB medium. Growth curve graphs – CFT073, MG1655 and derivative strains (.pdf): growth curve of CFT073, MG1655 and derivative strains in LB medium graphic representation. Growth curve graphs – EC071 and derivative strains (.pdf): growth curve of EC071 and derivative strains in LB medium graphic representation. Murine sepsis model – survival graph (.pdf): graphic representation of animal survival in a EC071 and derivative strains induced sepsis model. Murine sepsis model – survival statistics (.pdf): statistical analysis of animal survival in a EC071 and derivative strains induced sepsis model. pCF2 insertion in EC071 sat confirmation (.pdf): alignment of sequenced PCR products for pCF2 interruption of the sat gene in EC071 chromosome. pCF4 site-directed mutagenesis confirmation (.pdf): alignment of sequenced PCR products for nucleotide alterations in pCF4 sat gene. Proteolytic assays Immunoblotting (.pdf): detection of cleavage products after Sat and complement targets incubation. Sat or Sat’ production by pCF3 and pCF4 harboring strains (.pdf): detection of Sat ou Sat’ in TCA-treated culture supernatants of pCF3 and pCF4 harboring strains by SDS-PAGE, followed by silver nitrate staining and Immunoblotting. Sat production by EC071 and derivative strains (.pdf): detection of Sat in TCA-treated culture supernatants of EC071 and derivative strains by SDS-PAGE, followed by silver nitrate staining and Immunoblotting. Sat purification – mass spectrometry (.pdf): mass spectrometry analysis report for Sat identification in post-purification fractions. Sat purification – Silver staining and Immunoblotting: Sat detection in post-purification fractions by SDS-PAGE, followed by silver nitrate staining and Immunoblotting.