AatA plays a role in adhesion to chicken fibroblast DF-1 cells. A: Adhesion of AAEC189(pUC18) and AAEC189(pUC18: aatA
), expressing aatA under its native promoter, to DF-1 cells. Monolayers of DF-1 cells were incubated with E. coli strains for 3 h at 37°C. Adherent bacterial cells were harvested and the number was determined. B: The anti-AatA antibody inhibits binding capacity of IMT5155 to DF-1 cells. IMT5155 was incubated with preimmune serum (control) and with anti-AatA antibody, respectively. After washing, bacteria of each experiment were added to monolayers of DF-1 cells and incubated for 3 hours. Adherent bacterial cells were harvested and the number was determined. C: Pre-incubation of DF-1 cells with AatAF protein reduces adhesion capacity of IMT5155 to these cells. Confluent monolayers of DF-1 cells were incubated with BSA (control, 50 μg/well) or purified and refolded AatAF protein (50 μg/well) for 1 h at 37°C prior to the addition of IMT5155 cells. After 3 h of incubation, adherent bacterial cells were harvested and the number was determined. A-C: Columns represent the mean value of three independent wells per strain. Standard errors of the mean values are indicated as error bars. The experiment was repeated three times showing comparable results.