Comparison of membrane integrated and circulating ICAM-1 in wild-type and ICAM-1–exon 4 mutant mice.
(A) Visualization of the ICAM-1 expression on leukocytes isolated from wild-type and ICAM-1–exon 4 mutant mice. Isolated spleen cells were stimulated with 100 U/mL IFN-γ and 10 ng/mL LPS for 18 hours to increase ICAM-1 expression. Cells were incubated with FITC-conjugated anti-CD45 (1:7) and PE-conjugated anti-ICAM-1 (1:7). In each measurement PE-fluorescence intensity of 10 000 FITC-positive cells was determined. (B) cICAM-1 production by in vitro cultured spleen cells. Freshly isolated spleen cells (5 × 105 cells/well) were cultured for 3 days in RPMI supplemented with 10% FCS and 0.08% β-mercaptoethanol. Supernatant (150 μL) was taken after 1 hour and at the end of cultivation. cICAM-1 concentrations were determined by ELISA-2 in duplicate.
