FIGURE 2.

Inhibition of TREM-1 attenuates eCIRP-induced ICAM-1 expression on neutrophils in vitro. A, B, A total of 1 × 10⁶ BMDN isolated from WT and TREM-1^−/− mice were stimulated with PBS or rmCIRP at a dose of 1 μg/mL for 4 hours. After rmCIRP stimulation, BMDN were washed with PBS and stained with APC-Ly6G and FITC-ICAM-1 Ab. The frequencies of ICAM-1⁺ cells in LY6G gated population were assessed by flow cytometry. Experiments were repeated at least three times. Data are expressed as means ± SE (n = 5 samples/group). The groups were compared by one-way ANOVA and SNK method (*P < .05 vs PBS-treated WT BMDN; ^#P < .05 vs rmCIRP-treated WT BMDN). C, D, BMDN (1 × 10⁶ cells/ml) were treated with PBS, vehicle (PBS as LP17’s solvent), LP17 (80 μg/mL), and scramble peptide (80 μg/mL). After 30 minutes of the pretreatment of the cells with vehicle, LP17, and scramble, the cells were then stimulated with rmCIRP at a dose of 1 μg/mL for 4 hours. After stimulation with rmCIRP, the cells were washed with PBS and stained with APC-Ly6G and FITC-ICAM-1 Abs. ICAM-1 expression in LY6G⁺ cells were determined by flow cytometry. Experiments were repeated at least three times. Data are expressed as mean ± SE (n = 11 samples in PBS, vehicle, and LP17 groups; n = 6 samples in scramble group). The groups were compared by one-way ANOVA and SNK method (*P < .05 vs PBS-treated BMDN; ^#P < .05 vs vehicle + rmCIRP-treated BMDN; †P < .05 vs scramble + rmCIRP-treated BMDN)