Raw264

Cell lines used in this study were purchased from American Type Culture Collection unless otherwise specified. Murine macrophage cell lines RAW264.7 and J774A.1 were maintained in Dulbecco’s modified Eagle’s medium (DMEM high and low glucose, respectively; Biowest, France) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Life Technologies), 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin in a humidified incubator at 37°C under 5% CO₂. THP1 cells were maintained in RPMI-1640 (Biowest) with the supplements described above and THP1-XBlue-defMyD cells were purchased from InvivoGen and cultured according to the manufacturer’s instructions.

Murine macrophages (RAW 264.7) (ATCC-TIB-71) were acquired from the American Type Culture Collection (ATCC) biobank. RAW 264.7 cells were grown in DMEM (Dulbecco’s Modified Eagle’s Medium) without phenol red, supplemented with 10% FBS, 1% antibiotic/antimycotic solution (Biowest, Riverside, MO, USA), and 1% sodium pyruvate (humidified atmosphere with 5% CO₂ at 37 °C). Cells were seeded in 96-well microplates (5 × 10⁴ cells/well) for cell viability and nitric oxide (NO) production assessment. For determination of interleukins’ levels, the cells were seeded in 12-well microplates (5 × 10⁵ cells/well).