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For analyses of surface expression of adhesins by flow cytometry, 6 × 108 bacteria were washed once in PBS and then fixed for 20 min in 3% paraformaldehyde. Afterwards, bacteria were stained with the respective specific antiserum diluted 1:250 and a goat anti rabbit IgG antibody coupled to Alexa-Fluor488. Bacteria were measured with a FACSCalibur (Becton Dickinson) and analyzed using FCS Express (De Novo Software). A mutant strain devoid of the respective adhesin was always used as a negative control for gating.
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