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Expressing hPGAP1 and cPGAP1 (bottom) in HEK293 hPGAP1-KO cells (KO) restores PI-PLC sensitivity of CD59 staining in the FACS assay. Cells treated with (orange) or without PI-PLC (cyan) were analyzed by flow cytometry using gating strategies provided in Fig. S12a . PIGK-KO cells, in which GPI-AP biosynthesis are blocked 46 , 57 , are used as a staining control (gray). In our experiments ( n = 3 independent experiments), hPGAP1 reproducibly showed less activity than cPGAP1.

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