The MMTV-PyMT murine breast cancer model was prepared as described previously (9). A total of 5 × 105 MMTV-PyMT cells suspended in 50 μl of PBS were injected subcutaneously into the mammary gland on the right side of each mouse. Mice were treated with aldoxorubicin, Dox-SA, or Dox-CBD-SA on day 7 (5 mg/kg) via tail vein injection. Tumors were measured with a digital caliper at the indicated time points, and volumes were calculated as ellipsoids, where V = 4/3 × 3.14 × depth/2 × width/2 × height/2. Mice were euthanized when tumor volume had reached more than 500 mm3 or when active ulceration was observed. For the therapeutic experiment, FVB mice that originated from Charles River were used. For the TIL analysis, FVB mice that originated from both the Jackson Laboratory and Charles River were used. The proportion of mice from different providers was equalized among all groups.

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