LPS-induction of endothelial HA modification, but not neutrophil CD44 activity. (A) Flow cytometric analysis of FL-HA binding by mouse BM neutrophils left untreated, treated for 4 h with LPS (LPS) ex vivo, or treated with CD44-activating antibody IRAWB14.4 expressed as mean fluorescence intensity (MFI). (B) Adhesion of untreated (UT) and LPS-treated mouse neutrophils on coverslips coated with HA or P-selectin (P-sel; positive control) substratum in an in vitro flow chamber assay. Flow cytometric analysis of FL-HA binding by peripheral blood neutrophils left untreated (UT), treated with LPS ex vivo, incubated with IRAWB14.4, as well as neutrophils from LPS-treated mice (LPS in vivo) expressed as (C) a representative histogram plot and (D) as MFI. All results are representative of at least three experiments. Data are presented as the arithmetic mean ± SEM. *, P < 0.05 relative to untreated controls (on HA substratum for B). Livers of untreated (No LPS) and LPS-treated mice were visualized using spinning disk confocal microscopy. (E) The liver architecture shown by autofluorescence before the addition of any fluorophores and (F) SHAP localization observed by the administration of Alexa Fluor 488–labeled anti-SHAP mAbs are visualized. PE-labeled anti–Gr-1 mAb was used to demonstrate neutrophils adherent within sinusoids (E and F). Arrows denote locations of sinusoids. Bars, 50 μm. Images are representative of at least three experiments.