Gibco rpmi 1640

Human breast carcinoma cells HMC-1-8 (obtained from Creative Bioarray, Shirley, NY) and AU565^{50 (link)} were cultured in GIBCO^® RPMI 1640 (Thermo Fisher Scientific, VIC, AU) medium supplemented with 10% FBS (Thermo Fisher Scientific, VIC, AU) and 6 mM L-glutamine (Thermo Fisher Scientific, VIC, AU) at 37 °C with 5% CO₂. The identity of AU565 was confirmed through short tandem repeat profiling by CellBank Australia (Westmead, NSW, AU). Vector-, and TPD52-transfected stable BALB/c 3T3 cell lines have been previously reported^{39 (link)}, and were cultured in GIBCO^® RPMI 1640 medium supplemented with 10% FBS and 6 mM L-glutamine, with the addition of 1 mg/mL G418 (Geneticin®, Thermo Fisher Scientific, VIC, AU).

Most cell lines
were originally obtained from ATCC or internal stock. All cells were
genotyped to confirm their identity at the University of Michigan
Sequencing Core and tested routinely for Mycoplasma contamination.
SEM was grown in Gibco RPMI-1640 Glutamax + 10% FBS (ThermoFisher)
+ 1% Gibco MEM NEAA (100 × ) + 1% Gibco sodium pyruvate (100
mM) + 0.1% 2-mercaptoethanol (Sigma-Aldrich). RPMI-8402 was grown
in Gibco RPMI-1640 + 10% FBS (ThermoFisher). VCaP was grown in Gibco
DMEM + 10% FBS (ThermoFisher). 22RV1 was grown in Gibco RPMI-1640
+ 10% FBS (ThermoFisher). Human peripheral blood mononuclear cells
were obtained from Lonza. Sources of all antibodies and compounds
are listed in SI, Tables S1 and S2.

VR-2332, a parental virus of MLV (Ingelvac^® PRRS MLV, Boehringer Ingelheim, St. Joseph, MO, USA), which is the most commonly used vaccine in Korea, and two type 2 Korean PRRSV strains, K08-1054 (Accession number: JQ656266) and K07-2273 (Accession number: JQ656251), were used in this study. Based on the ORF5 amino acid sequence, K08-1054 and K07-2273 share 85.1% identity, and FL12 and K08-1054 or K07-2273 share 91% or 88.1% identity, respectively. VR-2332 shares a high sequence identity (96.5%) with K08-1054 and a low identity (84.6%) with K07-2273, whereas FL12 and VR-2332 share 90.5% sequence identity (Table 1). MARC-145, an African green monkey kidney cell line that is highly permissive to PRRSV, was used for virus propagation. MARC-145 cells were maintained in an Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco^® RPMI 1640, Life Technologies, Carlsbad, CA, USA) supplemented with heat-inactivated 10% fetal bovine serum (FBS, Life Technologies), 2 mM l-glutamine, and Antibiotic-Antimycotic 100× (Anti-anti, Life Technologies) containing 100 IU/mL penicillin, 100 µg/mL streptomycin, and 0.25 µg/mL Fungizone^® (amphotericin B). In this paper, this medium will be referred to as complete RPMI (cRPMI) medium. The cells were maintained at 37 °C in a humidified 5% CO₂ incubator.