Representative DEAE-sepharose fractionation of EA.hy926 endothelial cell membrane extracts to identify fractions to which <i>L. interrogans</i> bound.

<p>Top panel: FPLC system traces. The red trace denotes absorbance at 254 nm, the blue trace denotes absorbance at 280 nm, the brown trace denotes conductivity. Bottom panel: Binding of <i>L. interrogans</i> to fractions shown in the top panel. The “extract” was the octylglucoside supernatant loaded on to the column. All fractions were diluted 1∶10 in HBSC buffer without detergent prior to dispensing into the wells. After blocking the wells, 3.5×10⁵³⁵S-labeled pathogenic <i>L. interrogans</i> serovar Copenhageni was added to wells, and the plate was incubated 1 hour at 37°C, 5% CO₂ prior to removal of non-attached bacteria by washing. Buffer with octylglycoside, without or with NaCl, was used as a control for the start and end of the NaCl gradient, respectively.