Figure 3

Restoration of CD59 surface expression after PB-mediated rescue of PGAP2. (A) Schematic view of PGAP2 rescue using the PB transposon system. Plasmids containing PGAP2 and puroΔTK flanked by PB and FRT at both ends were transfected into cells together with a plasmid expressing PBase. After puromycin selection, cells stably expressing PGAP2 were obtained. (B) Restoration of surface expression in PGAP2-KO cells. PGAP2-KO3 cells were co-transfected with pCMV-hyPBase expressing PB transposase and pPB-FRT-PuroΔTK-PGAP2 to integrate the PGAP2 gene into the genome. CD59 on the cell surface was detected by flow cytometry. (C,D) After genomic DNA was extracted from P15 cells, the PB insertion site and PB copy number were determined in the genome. In P15 cells, only one PB copy band was detected (C). The full image of the gel was included in the supplementary information. The PB element was inserted into the intron region of the SBF2 gene region on chromosome 11 (D).