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Fig. 5. CA125-C-TERM cell-surface expression in CHOclone 13 cells does not result in increased binding capacity for exogenously added galectin-1. CA125-C-TERM-expressing and CA125-C-TERM-deficient CHOMCAT-TAM2 and CHOclone 13 cells were grown to 70% confluency. Where indicated, cells were treated with 10 µg/ml tunicamycin for 18 hours at 37°C. Cells were then dissociated from the culture plates followed by incubation with 40 µg/ml recombinant GST—galectin-1 for 30 minutes at room temperature. Following labeling with affinity-purified anti-galectin-1 antibodies under native conditions, the various samples were analyzed for cell-surface-bound recombinant galectin-1 using FACS. Autofluorescence (filled light-blue curve: CHOMCAT-TAM2; filled gray curve: CHOclone 13) was determined based on cells not treated with antibodies. Untreated CHOMCAT-TAM2 cells are shown in dark blue. Tunicamycin-treated CHOMCAT-TAM2 cells are shown in red. CHOclone 13 cells are shown in dark green (CA125-C-TERM-expressing) and light green (CA125-C-TERM-deficient), respectively.





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