FIGURE 9:

Gal-8 overexpression promotes anchorage-independent growth in soft agar and tumor generation in immunodeficient NSG mice. (A) Anchored-independent growth. MDCK-Gal-8^H cells (500 cells) seeded in soft agar generated an increased number of colonies, and larger, than MDCK cells. Analysis of 13 fields for each condition (number of colonies/field, mean ± SEM; **p < 0.005; two tailed unpaired t test with Welch´s correction. Areas of colonies from three experiments were measured with ImageJ software in 53 colonies of MDCK cells and 88 colonies of MDCK-Gal-8^H cells (mean ± SEM; *p < 0.05 two-tailed unpaired t test with Welch´s correction). Scale bar, 500 μm. (B) Tumor generation in NSG mice. Cells (10⁶) were subcutaneously injected in NSG mice, and the size of tumors was periodically assessed during 10 wk. The size of the nodules formed by MDCK cells remained similar from 2 to 10 wk, measuring around 1–1.5 mm in the killed animals (black arrow), whereas MDCK-Gal-8^H cells formed tumors in the range of 3–9 mm (longest diameter: scale bar, 5 mm). The tumor depicted in the figure measured around 9 mm and shows a region infiltrating the muscle (white arrow). Graph shows each tumor volume (**p < 0.005; Mann-Whitney test, two tailed).