Melanoma lung metastasis model

The B16-F10 melanoma cell line was obtained from ATCC (passage 3), and the experimental metastasis assay was performed as previously described (37, 38). C57BL/6 wild type mice, hCD1dKI, or VαKI mice were injected i.v. by the tail vein with 5 x 10⁵ B16-F10 cells in 200 µL of PBS. After 3 days, 4 nmol of glycolipids were administered by single i.v. injection in 0.2 ml vehicle (PBS + 0.05% Tween-20 + 0.1% DMSO). Two weeks after challenge, mice were sacrificed, lungs removed, and the area of melanized nodules on the lung surface was calculated from digital photographs of the excised lungs using Image J software from NIH (https://imagej.nih.gov/ij/index.html). Results were expressed as percentage of total lung surface area covered by melanized tumor growth.