FIGURE 2.

Genotyping and characterization of Fut8 disrupted cell clones. (A) Flow cytometry analysis of transfected CHO cells with Fu8‐KO donor cassette. NC represents untransfected CHO cells as negative control. (B) The fluorescence picture of EGFP positive cell clones after FACS sorting, the scale bar is 100 µm. (C) Genotyping analysis of ten EGFP positive and blasticidin survived cell clones. The length of wild type and knockout PCR products is 1.6 and 4.3 kb, separately. Het represents mono‐allelic mutation and homo represents biallelic mutation. (D) Flow cytometry analysis of LCA activity of fut8‐ko cell clones. The red histogram represents the wild type CHO cells without LCA treatments. Fut8(+/+) represents wild type CHO cells with LCA treatment for 30 min, Fut8(+/−) and Fut8(−/−) represents the fut8‐ko cells with LCA treatment for 30 min. The mean APC channel value of wild type CHO versus Fut8‐KO cells is 35000:320. (E) The morphology of wild type and fut8‐ko cells, the scale bar is 100 µm. (F) Cell growth of wild type and fu8‐ko cells. The cell numbers were calculated every two days during each subculture and identical numbers of cells were seeded for continuous passaging. C5 and C6 represent two cell clones with Fut8 mutation