Cell-bound galectin-3 does not potentiate PLM-A-induced TNF-α production.

<p>(<b>A</b>). The levels of endogenous galectin-3 bound to the J774 cell surface were determined by immunofluorescent staining using anti-galectin-3 antibody and FITC-conjugated secondary antibody. Cells were then fixed and analyzed by fluorescence microscopy. (<b>B</b>). Alternatively, cells were incubated with 10 µg/ml of exogenous rGal-3 for 1h at 20°C before staining and analysis performed as described in (A). Insert shows high magnification of labeled cells. (<b>C</b> and <b>D</b>) Cells were preincubated for 1h with (■) or not (□) 10 µg/ml of exogenous rGal-3 prior to the addition of 10 µg/ml of PLM-A or PLM-BMT6∆ (<b>C</b>) 10 µg/ml of curdlan or 500 ng/ml of Pam3CSK4 (<b>D</b>). Cells were cultivated for additional 4h. Then, supernatants were collected and TNF-α concentration was determined by ELISA. Results are presented as the mean ± standard deviation from 3 independent experiments. *<i>p<0</i>.<i>05; **p<0.01; ***p<0.0005</i>.