FIG 5.

AmOmpA interacts with α2,3-sialic acid and α1,3-fucose on mammalian host cell surfaces. RF/6A cells were pretreated with α2,3/6-sialidase (A to C), α1,3/4-fucosidase (A, D, and E), or vehicle control (A to E). Glycosidase- and vehicle-treated cells were incubated with α1,3/6-fucose-specific lectin, AAL (A); α2,3-sialic acid-specific lectin, MALII (A); His-AmOmpA (A to E); or media (cells alone; A to E). The cells were fixed and screened using immunofluorescence microscopy (A) or flow cytometry (B to E). In panel A, green fluorescence corresponds to lectin or His-AmOmpA bound at cell surfaces. Host cell nuclei are stained blue by 4′,6-diamidino-2-phenylindole (DAPI). For flow-cytometric data (B to E), representative histograms showing His-AmOmpA binding to RF/6A cells are presented in panels B and D. Mean fluorescence intensities ± SD from triplicate samples are presented in panels C and E. Data shown are representative of three independent experiments with similar results. Statistically significant (***, P < 0.001) values are indicated.