All bacterial strains and plasmids used in this study are listed in Table S1 . EHEC and E. coli HS were routinely grown in Luria-Bertani (LB) medium. E. faecalis V583 was routinely grown in Brain Heart Infusion (BHI) medium. B. thetaiotaomicron was routinely grown in TYG medium (41 (link)). For coculture experiments, Dulbecco’s modified Eagle’s medium with 1 g/L glucose (low-glucose DMEM) was used, as these conditions are known to induce EHEC virulence gene expression (42 (link)). Anaerobic growth was performed using a GasPak EZ anaerobe container system (Becton, Dickinson). When adequate, media were supplemented with ampicillin (100 μg/mL), gentamicin (50 μg/mL), chloramphenicol (20 μg/mL), erythromycin (10 μg/mL), or streptomycin (100 μg/mL). Bacterial stocks were kept on LB supplemented with glycerol 30% (vol/vol) at −80°C.
HeLa (human cervical adenocarcinoma) cells were maintained in DMEM with 4.5 g/L glucose (high-glucose DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin-glutamine. The Lifeact::GFP-expressing HeLa cell line was obtained with the Flip-In system (Invitrogen) as previously described (43 (link)), and maintained in high-glucose DMEM supplemented with 10% FBS, penicillin-streptomycin-glutamine, and hygromycin (50 μg/mL). Hygromycin was not added when cells were split before infection. Cells were routinely grown at 37°C and 5% CO2.
HeLa (human cervical adenocarcinoma) cells were maintained in DMEM with 4.5 g/L glucose (high-glucose DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin-glutamine. The Lifeact::GFP-expressing HeLa cell line was obtained with the Flip-In system (Invitrogen) as previously described (43 (link)), and maintained in high-glucose DMEM supplemented with 10% FBS, penicillin-streptomycin-glutamine, and hygromycin (50 μg/mL). Hygromycin was not added when cells were split before infection. Cells were routinely grown at 37°C and 5% CO2.
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