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. 2009 Nov 17;285(4):2232–2244. doi: 10.1074/jbc.M109.066191

FIGURE 3.

FIGURE 3.

Expression of core 2 O-glycans increases galectin-1 binding and is required for galectin-1 inhibition of CD45 tyrosine phosphatase activity in cells expressing CD45RB. A, expression of C2GnT adds core 2 O-glycans to T cell surface glycoproteins. PhaR2.1, BW5147, BW-C2GnT, or BW-vector cells were stained with CD45-FITC or 1B11-PE (open) or isotype control (filled). 1B11 binding to BW-C2GnT cells demonstrates core 2 O-glycan addition. B, BW-C2GnT (squares) or BW-vector (circles) cells were bound by biotinylated galectin-1 for 10 min with (open) or without (filled) 50 mm lactose. Bound galectin-1 was detected with streptavidin-FITC, and MFI was determined. Data are mean fluorescence, corrected for autofluorescence, ± S.D. of duplicate samples. C, 1 × 106 PhaR2.1, BW-vector, or BW-C2GnT cells were incubated with 20 μm galectin-1 (circles) or buffer control (squares) for the shown times. Cells were then washed and lysed, and phosphatase activity was measured with (open) or without (filled) bpV(phen). Data are mean ± S.D. of triplicate samples.