Bone marrow isolation from female mice (age 6-8 weeks, FVB/Hsd strain from Envigo) was performed in accordance with the Italian Laws (D.L.vo 116/92 and following additions), which enforce the EU 86/609 Directive. All animal procedures were approved by the OPBA (Organismo per il Benessere e Protezione Animale) of the Cogentech animal facility at the IFOM-IEO Campus, Milan. Bone marrow derived macrophage (BMDM) cultures were carried out as described61 (link). LPS from E.Coli serotype 055:B5 (cat. no. L4524, Sigma) was used at 10 ng/ml.
HeLa and HEK 293 cells (both from ATCC) were cultured in DMEM with South American serum (10%), Pen/Strep (1%, cat. no. P4333, Sigma) and L-Glutamax (1%, cat. no. 35050061, Gibco). RAW264.7 cells (from ATCC) were cultured in DMEM with North American serum (10%), Pen/Strep (1%) and L-Glutamax (1%). Cell lines were authenticated by the Tissue Culture Facility of IEO using the GenePrint10 System (Promega) and routinely screened for Mycoplasma contamination.
HeLa and HEK 293 cells (both from ATCC) were cultured in DMEM with South American serum (10%), Pen/Strep (1%, cat. no. P4333, Sigma) and L-Glutamax (1%, cat. no. 35050061, Gibco). RAW264.7 cells (from ATCC) were cultured in DMEM with North American serum (10%), Pen/Strep (1%) and L-Glutamax (1%). Cell lines were authenticated by the Tissue Culture Facility of IEO using the GenePrint10 System (Promega) and routinely screened for Mycoplasma contamination.