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Cytolytic Assay: Gating Strategy

LH Leen Hermans
SP Steffi De Pelsmaeker
SD Sofie Denaeghel
EC Eric Cox
HF Herman W. Favoreel
BD Bert Devriendt
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Samples were analyzed using a Cytoflex (Beckman Coulter). Doublet discrimination was carried out based on FSC-H/FSC-A and SSC-H/SSC-A and cells were selected based on their FSC-A/SSC-A properties. CFSE-labeled target cells were selected on FSC-A/FITC-A, after which CFSE-positive cells were gated on cell viability on FSC-A/PC5.5-A. For all conditions 5,000 CFSE-labeled target cells were measured and the percentage of NK mediated lysis was calculated using the formula (% dead targetNK – % dead targetspont)/(%dead targetmax – % dead targetspont).

Copyright and License information:  ©2021 Hermans, De Pelsmaeker, Denaeghel, Cox, Favoreel and Devriendt.
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