V. cholerae strains were grown at 30°C in LB medium supplemented with streptomycin (100 μg ml−1). Overnight cultures were diluted 1:100 and grown at 30°C with shaking until exponential phase [OD600 (optical density at 600 nm) ≈ 0.40 to 0.60]. Bacteria from 1 ml were pelleted by centrifugation and resuspended in PBS to a final concentration of 5 × 108 bacterial cells/ml. T84 cells (seeded the day before into cell culture–treated plates) were twice washed with PBS, and then, media were changed to antibiotic-free and FBS-free DMEM/F12. Resuspended V. cholerae was added to the media over cells (MOI = 5). Inoculations were synchronized by centrifugation at 500g for 3 min. T84 cells were subsequently incubated at 37°C in the presence of 5% CO2 until processed for downstream applications.

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