Investigating CD73 in 4T1 Breast Cancer

Four-week-old female Balb/c mice (Balb/cOlaHsd) were obtained from Envigo (Netherlands). Animals were maintained under controlled pathogen-free environmental conditions with a 12h light/dark cycle. Mice were inoculated with sh-NT and sh-CD73 4T1cells (2×10⁴ cells in 100 μl PBS per mouse) orthotopically into 4^th mammary fat pads (n = 10/group) and followed for 31 days. For the metastasis models, the mice were inoculated intravenously with sh-NT and sh-CD73 4T1 cells (5×10⁴ cells in 100 μl PBS per mouse) into tail vein (n = 6/group) and followed for 20 days. In the B cell depletion model, 100 µM/animal Ultra-leaf purified anti-mouse CD20 (BioLegend, 152104) and control IgG antibody (BioLegend, 400671) were injected intravenously in the tail vain, once cells were inoculated and followed for 34 days. Animals were treated intraperitoneally each 4^th day with the dose of 6 µg ZOL/animal. Body weights and tumor dimensions (35 (link)) were measured once a week. The animals were sacrificed when weight loss was ≥ 10% (data not shown).