Fig. 5. CA125-C-TERM cell-surface expression in CHO_{clone 13} cells does not result in increased binding capacity for exogenously added galectin-1. CA125-C-TERM-expressing and CA125-C-TERM-deficient CHO_MCAT-TAM2 and CHO_{clone 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: CHO_MCAT-TAM2; filled gray curve: CHO_{clone 13}) was determined based on cells not treated with antibodies. Untreated CHO_MCAT-TAM2 cells are shown in dark blue. Tunicamycin-treated CHO_MCAT-TAM2 cells are shown in red. CHO_{clone 13} cells are shown in dark green (CA125-C-TERM-expressing) and light green (CA125-C-TERM-deficient), respectively.