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Establishment of a robust in vitro PIG-A assay gating strategy. Utilising viability dye incorporation (Annexin V and 7-AAD) and tandem markers (CD55 and CD59) the proposed robust reproducible gating strategy (B) was directly compared against traditional flow cytometry gating (A). Retrospectively, Day 3, 24 h MNU (0–1 µg/mL) exposure data within MCL-5 cells was assessed using both methods in order to estimate phenotypic PIG-A mutant frequency and hence, dose response.
