PMC:7196351 / 10719-11435 JSONTXT

{"project":"2_test","denotations":[{"id":"32373300-20603831-54158470","span":{"begin":239,"end":241},"obj":"20603831"},{"id":"T95078","span":{"begin":239,"end":241},"obj":"20603831"}],"text":"To discriminate necrotic/late apoptotic from early apoptotic cells, the presence of PS on the outer surface of the apoptotic cells was detected from fluorescein isothiocyanate (FITC)-conjugated annexin V binding to PS at the cell surface (14), and necrosis/late apoptosis was assessed from the amount of propidium iodide (PI)-positive cells. Briefly, 106 splenic leukocytes were harvested and washed twice with annexin washing buffer (AWB, 10 mM Hepes/NaOH, pH 7.4, 140 mM NaCl, 5 mM CaCl2). The cell pellet was resuspended in 100 µl of annexin-V/PI labelling solution (eBioscience, USA), and incubated for 15 min at room temperature. After washing with AWB, CD45-positive lymphocytes were analysed by flowcytometry."}