FIG. 6.

(A and B) SDS-PAGE analysis of purified FliC obtained from the wild-type strain (panel A) and Western blot with anti-His and anti-flagellin antibodies (panel B, lanes 1 and 2, respectively). (C) HBMEC were preincubated with 100 μg of the preparation or 10 mM Tris-HCl for 1 h, followed by an association assay with the wild type or the fliC mutant with centrifugation. Preincubation treatments are indicated in parentheses. One hundred micrograms of His-tagged AslA was used as a nonspecific His-tagged protein control. Preincubation with FliC significantly (P < 0.05) blocked association of the wild type but had no effect on the fliC mutant. The bars indicate the means of two independent experiments, and the error bars indicate the standard deviations.