Figure 1.

Effect of monocyte viability on BzATP-stimulated P2X₇ pore function. A. Peripheral blood was obtained from eight laboratory volunteers and processed immediately or stored at room temperature for up four days to simulate the conditions of shipping blood from centers around the country. On the day of processing, aliquots of the blood samples were stained and data was acquired using bead-adjusted settings on a FACSCalibur flow cytometer. Data is expressed as % CD14^pos/PI^neg events in the sample (mean ± SEM; n =8). B. Anti-CD14-PE labeled human periphral blood from a representative subject was stimulated with control vehicle or 250 mM BzATP for 20 minutes in the presence of 1 mM YO-PRO-1 dye as described in Materials and Methods and beads were used to set the cytometer. Overlayed histogram analysis of YO-PRO-1 flurescence detected in live (dashed or solid outline) and total (light or dark shading) monocyte populations with or without stimulation with BzATP as denoted by the inset symbol key. The arrow indicates the presence of non-viable monocytes in both the saline and the BzATP samples when analyzing total events, that are also separate populations on PE vs YO-PRO-1 plots.