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Fig. 2 (A and B) Flow-cytometry histograms of FucT-treated (red) and ISF-treated (blue) Pro-5 CHO cells after 90 min of incubation. LRP6 in (B) was transiently-expressed FLAG-LRP6. (A) Detection of cell surface N-glycan LacNAc fucosylation by the lectin AAL-FITC. (B) Immunofluorescent staining of total cellular LRP6 and cell-surface LRP6. (C) Scheme of LRP6 location and maintenance regulated by endocytosis. (D) Ratio of LRP6 (surface to total) in Pro-5 cells treated by ISF or FucT only. Mock control was set as 100%. Error bars show standard error. N = 53,664 cells. Unpaired two-tailed Student's t test: ∗∗∗∗p < 0.001. (E) Western blot assay of membrane LRP6, lipid raft LRP6, cytoplasmic LRP6, and actin after different times of ISF treatment. Number indicates the relative LRP6 level normalized to the corresponding level of actin.