Figure 6
Enrichment of cells highly expressing LIPA. (A) Sorting of cells highly expressing sHF-LIPA-GPI. P15 cells stably expressing sHF-LIPA-GPI were sorted twice. Surface expression of sHF-LIPA-GPI was analysed by flow cytometry. Cells were stained with an anti-Flag antibody followed by a PE-conjugated goat anti-mouse IgG. (B) Sorted cells were transfected with the vector carrying PBase or FLPe recombinase. Expression of CD59 and sHF-LIPA-GPI on the cell surface was analysed. Original, P15 cells expressing sHF-LIPA-GPI after one round of sorting (Sort 1) and after two rounds of sorting (Sort 2). (C) sHF-LIPA in the cell lysate and culture medium was analysed. Culture medium from Sort 1 and Sort 2 cells with or without (−) treatment of PBase or FLPe recombinase were collected and precipitated. Proteins were detected by anti-Flag antibodies. The full image was included in the supplementary information.
