Clone rb6 8c5

For cell depletion studies, mice were randomized at 50mm³ tumor volume and injected with 100 μL antibodies through the intraperitoneal route. An initial depletion with 400 μg of anti-CD8a antibody (Cat: BP0061, clone 2.43, BioXcell) or rat IgG2b isotype control (Cat: BE0090, BioXcell) was administered on days 7, 11, and 15 post-tumor inoculation. Antibody depletion treatment was continued with the administration of 400 μg of antibody every 3-5 days for the duration of the experiment. For depletion of MDSCs, ~1w post tumor inoculation, mice with palpable tumors were injected intraperitoneally (IP) with anti-GR1 antibody 200 μg/dose (Cat: BE0075, clone RB6-8C5, BioXcell) or isotype IgG2b (Cat: BE0090, BioXcell) every 48 hours. On Day 18 after the first anti-GR1 antibody administration, the lungs were harvested to evaluate for metastasis. Tumors & other organs were also harvested to confirm the depletion of MDSC. Depletion efficiency was checked using FACS analysis of blood obtained by retro-orbital bleeding. For conditioned media treatment, concentrated CM from WT-Gal1 or KO-Gal1 cells or similarly concentrated control media was used. When tumors reached ~100mm³ in size, 100μl of CM/control media was injected IP every other day for ~3 weeks until termination.