E0771

The 4T1 and E0771 tumor cell lines were purchased from the American Type Culture Collection (ATCC) and CH3 BioSystems, respectively. The AT-3 cell line was gift from Dr. Scott Abrams (Roswell Park). Tumor cells expressing luciferase (4T1-luc, E0771-luc, and AT-3-luc) were generated with infection of lentiviruses encoding luciferase (pLenti PGK V5-LUC Neo, Addgene plasmid #21471). 4T1 and E0771 cells were cultured in RPMI 1640 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% Non-Essential Amino Acid (NEAA) (Gibco), 2 mM L-glutamine (Gibco), 0.5% penicillin/streptomycin (Gibco), and 55 μM 2-mercaptoethanol (Gibco). AT-3 and AT-3-luc cells were cultured in DMEM (Gibco) supplemented with 10% FBS, 1% NEAA, 2 mM L-glutamine, 0.5% penicillin/streptomycin, and 55 μM 2-mercaptoethanol. These cell lines were authenticated by morphology, phenotype and growth, and routinely screened for Mycoplasma, and were maintained at 37°C in a humidified 5% (4T1 and E0771) or 7% (AT-3) CO₂ atmosphere.

4T1 and YAC-1 cells were purchased from the ATCC. E0771 cells were purchased from CH3 Biosystems. AT3 cell line was a generous gift from S.I. Abrams at Roswell Park Comprehensive Cancer Center. To generate AT3-Luc, E0771-Luc or 4T1-Luc cells, the tumor cells were infected with luciferase-expressing lentivirus (Addgene) (Campeau et al., 2009 (link)). For OVA labeling, the AT3-Luc cells were transfected with pcDNA3-OVA (Addgene) (Diebold et al., 2001 (link)) to generate AT3-OVA-Luc tumor cells. For mCherry labeling, AT3 cells were infected with mCherry-expressing lentivirus (Addgene) to generate AT3-mCherry cells. To overexpress G-CSF, AT3 cells were infected with g-csf (Csf3)-expressing lentivirus (the vector was a gift from R.A. Weinberg, Massachusetts Institute of Technology) to generate AT3-gcsf cells. To generate 4T1-GFP, AT3-GFP, AT3-gcsf-GFP and E0771-GFP cells, the tumor cells were infected with green fluorescent protein (GFP)-expressing lentivirus (Addgene). Human lung fibroblast-like cells (mesenchymal stem cells) were purchased from ScienCell Research Laboratories. All cell lines were maintained in a humidified 5% CO₂ incubator at 37 °C and confirmed to be mycoplasma free.