Fig. 4.
Fig. 4. Kinetics of the effect of U46 619 and free AA on cytosolic Ca++ of erythrocytes. / RBCs loaded with fura-2 am (0.1% hematocrit in HEPES buffer) were placed in a quartz cuvette with stirring (37°C, 300 rpm) for fluorescence measurements (“Materials and methods”). Fluorescence was determined with excitation wavelengths 340 and 380 nm and 510 nm emission wavelength in a spectrofluorimeter. After determination of baseline [Ca++]i, U46 619 (1 μM), free AA (20 μM), ionomycin (1 μM), or solvent (controls) was added. [Ca++]i of erythrocytes was quantified at 1-second intervals for 10 minutes. The figure shows time-dependent [Ca++]i changes. Data are representative of 11 separate experiments in different donors. PGI2 (0.5-1 μM) or 12-HETE (0.1-1 μM) did not increase [Ca++]i over that of solvent control (not shown).

Kinetics of the effect of U46 619 and free AA on cytosolic Ca++ of erythrocytes.

RBCs loaded with fura-2 am (0.1% hematocrit in HEPES buffer) were placed in a quartz cuvette with stirring (37°C, 300 rpm) for fluorescence measurements (“Materials and methods”). Fluorescence was determined with excitation wavelengths 340 and 380 nm and 510 nm emission wavelength in a spectrofluorimeter. After determination of baseline [Ca++]i, U46 619 (1 μM), free AA (20 μM), ionomycin (1 μM), or solvent (controls) was added. [Ca++]i of erythrocytes was quantified at 1-second intervals for 10 minutes. The figure shows time-dependent [Ca++]i changes. Data are representative of 11 separate experiments in different donors. PGI2 (0.5-1 μM) or 12-HETE (0.1-1 μM) did not increase [Ca++]i over that of solvent control (not shown).

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