FIG 2 .

Repression of alternative glucose transport and biofilm formation is mediated by phospho-EI and relieved by EIIA^Glc AH-independent phosphotransfer to EIIB. (A and B) Phos-tag acrylamide gel electrophoresis and anti-V5 antibody immunoblotting of the affinity-tagged full-length EIIA^Glc (FL), full-length EIIA^Glc with histidine 91 mutated to alanine (FL^H91A), Δ16 EIIA^Glc (Δ16), Δ16 EIIA^Glc with histidine 91 mutated to alanine (Δ16^H91A), or EIIA^Glc in a strain with histidine 189 of EI mutated to alanine (EI^H189A) extracted from cells grown in LB alone or supplemented with glucose. (C and D) A₆₁₅ measured over time for V. cholerae expressing the indicated EIIA^Glc alleles or deletion (ΔEIIA) and cultured in minimal medium supplemented with pH indicators and glucose. (E and F) Quantification of biofilm formation by the indicated V. cholerae strains cultured in MM supplemented with glucose. Data represent the means from three biological replicates, and error bars represent the standard deviation. Statistical significance was calculated using a one-way analysis of variance followed by Tukey’s multiple-comparison test. ns, not significant. (G) A₆₁₅ measured over time in cultures of the indicated V. cholerae control or mutant strains. Minimal medium supplemented with pH indicators and glucose was used. Results are representative of three independent experiments. (H) Quantification of biofilm formation by the indicated V. cholerae strains cultured in MM supplemented with glucose. Data represent the means from three biological replicates, and error bars represent the standard deviation. Statistical significance was calculated using a one-way analysis of variance followed by Tukey’s multiple-comparison test.