Rpmi 1640 culture media

Reagents and antibodies were purchased as follows: Ficoll-Hypaque from GE Healthcare Bio-Sciences AB (Uppsala, Sweden), RPMI-1640 culture media and Fetal bovine serum (FBS) from Invitrogen (Carlsbad, CA, USA), Dulbecco’s Modified Eagle’s Medium (DMEM; 12,800,017) from Thermo Fisher (Carlsbad, CA, USA), PE conjugated mouse antibody directed to CD11b and isotype controls from BD Bioscience (San Jose, CA, USA), TSLP from R&D Systems (Minneapolis, MN, USA), TSLPR antibody from Biolegend (San Diego, CA, USA), Programed death-ligand 1 (PDL1) and REGF antibodies from Biolegend (San Diego, CA, USA), and TMZ Sigma (St. Louis, MO, USA).

LNCaP and PC-3 cell lines were obtained and maintained as described previously30 (link). Cisplatin and MG132 were purchased from Sigma (St. Louis, MO, USA), and. Methyltrienolone (R1881) was purchased from NEN life Science (Boston, MA, USA). The RPMI-1640 culture media were purchased from Life Technologies (Rockville, MD, USA). Fetal calf serum (FCS) was purchased from the HyClone (Logan, Utah, USA).

The SW480 and HCT116 CRC cell lines were purchased from the American Type Culture Collection (ATCC, USA). The cell lines were grown in Gibco Roswell Park Memorial Institute 1640 (RPMI-1640) culture media (Life Technologies, USA) supplemented with 10% heat-inactivated Gibco fetal bovine serum (FBS) (Life Technologies, USA) at 37 ºC with 5% carbon dioxide (CO₂) concentration. The adherent cell lines were detached using Gibco 0.05% trypsin-EDTA (Life Technologies, USA) dissociation agent, washed with 1 X PBS and centrifuged for 5 min at 100 X g. The supernatant was removed after centrifugation and the pellet was resuspended in 1 mL of media before cells counting.

Murine mesothelioma cell line AK7 was provided by Brown University, USA. RN5 and ZiP3 cell lines were developed from C57BL/6 background mice after intraperitoneal injection of asbestos fibers [25 (link),26 (link)]. The AE17 cell line was provided by Dr. Steven Albelda, University of Pennsylvania (Philadelphia, PA, USA), and Dr. Delia Nelson (University of Western Australia, Crawley, Australia). All cell lines were grown in RPMI1640 culture media (Life Technologies Inc. Burlington, ON, Canada) supplemented with 10% heat-inactivated FBS (Life Technologies Inc.), 2 mmol/L L-glutamine, 100 U/mL penicillin, 100 mg/mL streptomycin and nonessential amino acids. Cells were plated in tissue-culture coated flasks (BD Biosciences Canada, Mississauga, ON, Canada), grown in a 37 °C and 5% CO₂ environment, and passaged when 70% confluent. Eight to 12 weeks old C57BL/6 syngeneic mice and IRF3KO mice were initially purchased from Jackson Laboratories, and acclimatized in the animal colony for 1 week before experimentation. The animals were housed in microisolator cages, 5 per cage, in a 12 h light/dark cycle. Sterile water and rodent food were given ad libitum.