Figure 1.

Galectin-3 accumulation did not alter upon exposure of cells to hypoxic conditions. The normal murine melanocyte cell line, Melan-a, gave rise to the melanoma cell line Tm1, as described elsewhere (Oba-Shinjo et al. 2006). Lack of galectin-3 expression was among the differences between the tumor cell lines and their normal counterpart (de Souza et al. 2006). We had then transfected the galectin-3 gene in the tumorigenic cell line Tm1, obtaining clones such as the ones used in this study. Control transfectants were obtained by transfection of the empty plasmid pEF1.neo (Tm1.Neo cells, Tm1.Nx cells) and the galectin-3 expressing cell was obtained by transfection of the galectin-3 coding plasmid pEF1.neo/gal3 (Tm1.Gal-3, Tm1.Gx cells). Protein extracts of the cell lines studied were routinely analyzed by western blots. Loading controls were done analyzing β-actin accumulation and galectin-3 expression was analyzed with specific monoclonal antibodies (M3/38, rat anti-galectin-3). Exposure of cells to hypoxia (atmospheres containing 1% oxygen) for 24 hours did not interfere with accumulation of galectin-3, differently from the exposure of cells to cobaltous chloride, a prolyl-hydroxylase inhibitor (and so-called “hypoxic mimetic” drug). Western blots were done after resolving protein extracts in 10% SDS-polyacrylamide gels. Plus and minus signs represent whether cells were exposed or not to a given experimental condition.