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After incubation for 1 h at 37°C, either cholic acid (CA), glycocholic acid (GC), taurocholic acid (TC), deoxycholic acid (DOC), taurodeoxycholic acid (TD), glycodeoxycholic acid (GD), glycine (Gly), or taurine (Tau) were added to the mucin columns at a final concentration of 1.2 mM each. After incubation for 2 h at 37°C, cells were collected, serial dilutions were plated on LB agar plates, and surviving numbers of C6706 or C6706ΔtsiV1–3 were plotted. Bars show mean values ± SD of two independent experiments done in duplicate. (B) Individual bile acids affect the T6SS of V. cholerae V52. Predator V. cholerae V52 or V52ΔvasK were mixed at a 10:1 ratio with prey E. coli MG1655 and spotted on LB agar plates supplemented with 1.2 mM cholic acid (CA), glycocholic acid (GC), taurocholic acid (TC), deoxycholic acid (DOC), glycodeoxycholic acid (GD), taurodeoxycholic acid (TD), glycine (Gly), or taurine (Tau) for 4 h at 37°C. CFUs were counted after incubation of serial dilutions of eluent on LB agar plates overnight. The killing index was calculated by the ratio of surviving prey in the presence of V52 versus attenuated V52. The graph gives mean values ± SD of two experiments done in duplicate. 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