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Cells were isolated from blood and brain samples for flow cytometric analysis to identify GFP+ cells in the tissues.
Samples for flow cytometry were centrifuged for 1 min at 3000g and 4°C, and resuspended in saline solution.
Samples for flow cytometry were washed with PBS, resuspended in cryo-protective solution (15% (v/v) glycerol in PBS according to Jahn et al. ([2013])) and stored at −20°C.
Samples for flow cytometry and microscopy were taken at different time points after UV irradiation.
Preparation of the samples for flow cytometric analysis was performed as described previously [9], [10], [11].
Samples for flow cytometry and PCR were taken on days 0 28 of differentiation and analyzed as described below.
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We treated asynchronous cultures with 10 mM HU for 4 hours, released them in fresh media and collected sample for flow cytometry for every 20 minutes.
This apparently discrepant result may be due to the population of cells sampled for flow analysis not being representative of the population of cells with loss of A allelic expression, or the malignant cells not being able to differentiate to RBCs.
From metamorphs and juveniles we took a toe clip for microsatellite analysis and a blood sample for flow cytometry analysis.
Cultures were sampled for flow cytometry after induction with β-estradiol at T=2 hours and T=19 hours.
Cell counting and sampling for flow cytometry and microarray analysis were carried out at 0, 4, 10, 48 and 96 hours in the CD3+ T-cell experiments, E1-E5, with cells from 5 independent healthy donors, and at 0, 6, 12, 24, 48 and 72 hours in the CD4+ T-cell and CD8+ T-cell experiments, E7-E11, with cells from 5 independent healthy donors.
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