Mcherry expressing lentivirus

4T1, YAC-1, NK-92MI and K562 cell lines were all purchased from the American Type Culture Collection (ATCC). AT3 cell line was kindly provided by S.I. Abrams (Roswell Park Comprehensive Cancer Center). E0771 cell line was obtained from CH3 Biosystems. Human lung mesenchymal cells were purchased from ScienCell Research Laboratories. MDA-4175 cells, originally generated from J. Massagué’s laboratory (Memorial Sloan-Kettering Cancer Center), was kindly provided by Y. Kang (Princeton University). 4T1 and AT3 cell lines were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin YAC-1, K562 and E0771 cell lines were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin NK-92MI cell lines was cultured in the Alpha Minimum Essential medium supplemented with 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, 12.5% horse serum and 12.5% FBS. AT3 cells were infected with mCherry-expressing lentivirus (Addgene, plasmid #36084) to generate AT3-mCherry cells. For luciferase labeling, 4T1, AT3, E0771 and MDA-4175 cells were infected with luciferase-expressing lentivirus (Addgene, plasmid #17477) ^{93 (link)}. All cell lines used in this study were tested and confirmed to be negative for mycoplasma.

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.