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. 2018 Feb 5;217(2):585–599. doi: 10.1083/jcb.201706135

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© 2018 Liu et al.

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Figure 6. — Overexpression of misfolded GPI-APs decreased deacylation efficiency of endogenous GPI-APs in WT but not CANX-KO cells. (A) WT HEK293FF6 or CANX-KO cells stably expressing WT EGFP-F–CD59, mutant CD59 (C94S and misfolded CD59), misfolded CD59 lacking GPI (C94S and G103stop*), or misfolded CD59 lacking an N-glycan (C94S and N43Q) were treated with or without PIPLC. After treatments, cells were stained for endogenous DAF and analyzed by flow cytometry. (B) DAF levels remaining after PIPLC treatment were plotted. The value for remaining DAF in WT cells without any exogenous expression was set to 1. (C) WT HEK293FF6 or CANX-KO cells stably expressing WT EGFP-F–DAF or mutant DAF (C81A, misfolded DAF) were treated with or without PIPLC. After treatment, cells were stained for endogenous CD59 and analyzed by flow cytometry. (D) CD59 levels remaining after PIPLC treatment are shown. The remaining CD59 value in WT cells without any exogenous expression was set to 1. Relative values were calculated and are represented as means ± SD from three independent experiments.