Fig. 3.
Fig. 3. Cell adhesion molecule expression in HUVEC in response to interaction with RBC. HUVEC grown to confluence in 24-well plates were incubated with RBC (2% Hct) and E-CM (100 μg/mL) for 4-hour time periods, unless otherwise indicated. ICAM-1, E-selectin, and VCAM-1 expression was determined by ELISA. (A) Time period of expression of CAMs in response to SS RBC plus E-CM; results are expressed as mean ± SD of OD at 405 nm of triplicate determinations, (n = 5; including three different donors). (B) Effect of AA RBC (n = 3), AS RBC (n = 3), and SS RBC (n = 6; including four different donors) on CAMs expression at 4 hours. (C) HUVEC were preincubated for 30 minutes with peptides (KYRGDS, 100 μmol/L; AGDV, 100 μmol/L) and polymyxin sulfate (5 μg/mL) before incubation with SS RBC plus E-CM or E-CM (100 μg/mL) alone for 4 hours. Incubation with LPS (100 ng/mL) was performed for 4 hours. (D) HUVEC were preincubated for 30 minutes with inhibitors (Probucol, 50 μmol/L; vitamin E, 25 μmol/L; SOD, (200 U/mL); catalase, (200 U/mL); GSH-EE, 0.5 mmol/L; and BSO (100 μmol/L) before incubation with SS RBC for 4 hours, followed by measurement of VCAM-1 expression.

Cell adhesion molecule expression in HUVEC in response to interaction with RBC. HUVEC grown to confluence in 24-well plates were incubated with RBC (2% Hct) and E-CM (100 μg/mL) for 4-hour time periods, unless otherwise indicated. ICAM-1, E-selectin, and VCAM-1 expression was determined by ELISA. (A) Time period of expression of CAMs in response to SS RBC plus E-CM; results are expressed as mean ± SD of OD at 405 nm of triplicate determinations, (n = 5; including three different donors). (B) Effect of AA RBC (n = 3), AS RBC (n = 3), and SS RBC (n = 6; including four different donors) on CAMs expression at 4 hours. (C) HUVEC were preincubated for 30 minutes with peptides (KYRGDS, 100 μmol/L; AGDV, 100 μmol/L) and polymyxin sulfate (5 μg/mL) before incubation with SS RBC plus E-CM or E-CM (100 μg/mL) alone for 4 hours. Incubation with LPS (100 ng/mL) was performed for 4 hours. (D) HUVEC were preincubated for 30 minutes with inhibitors (Probucol, 50 μmol/L; vitamin E, 25 μmol/L; SOD, (200 U/mL); catalase, (200 U/mL); GSH-EE, 0.5 mmol/L; and BSO (100 μmol/L) before incubation with SS RBC for 4 hours, followed by measurement of VCAM-1 expression.

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