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Solubilized samples were centrifuged at 17,400 g for 30 min at 4 C, and 13.5 l of the supernatant was mixed with 1.5 l of 10 loading buffer (50% glycerol, 0.1% Ponceau S)

Interestingly, when we optimized the cell sorting procedure by using free settling to replace magnetic separation, the EGFP positive cell percentage could be elevated from 20% to over 95% in K-562 cells for both GST-EGFP-GPI DAF and GST-EGFP-GPI BY55 (Figure 3C)