To establish a bilateral 4T1 and TMT6 tumor model, female Balb/c mice (18-20 g) were injected subcutaneously with 1 × 106 4T1 or EMT6 cells in the left flank (primary tumor), one week or 6 days after inoculation of 4T1 or EMT6 respectively, 1 × 105 4T1 cells or 5 × 105 EMT6 cells were injected into the medullary cavity of the right tibia through a 26-gauge needle under anesthesia (bone metastatic tumor).
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