Murine 4T1 Breast Cancer Model

Tumor cell preparation was adopted from the literature.^{32 (link)} Briefly, 4T1 Luc cells were cultured at 37 °C with 5% CO₂ in a T75 flask (BD Biosciences, Bedford, MA); DMEM (Dulbecco’s Modified Eagle’s medium, Gibco, USA) medium supplemented with 10% FBS and 50 U/mL penicillin/streptomycin. After 3 passes, the cells were suspended in the DMEM and 1 × 10⁵ cells were injected subcutaneously on top of the right leg of female BALB/c mice (6–8 weeks old, body weight ~20 g). The mice were monitored for approximately two weeks post-inoculation until the tumor size was between 7–10 mm.