Hr csf 1 m csf

THP-1 cell line was obtained from ATCC (Cat#ATCC TIB-202) and maintained in RPMI 1640 (Corning, Cat#10-040-CV) supplemented with 10% FCS and 1% Pen/Strep and were differentiated into macrophages in the presence of 10 ng/ml phorbol-12-myristate acetate (PMA, Sigma, Cat#P8139) for 12–16 h followed by media change and resting for up to 48 h. CD14 + monocytes were differentiated into monocyte-derived Dendritic Cells (hMDDCs) using a cocktail of hIL-4 and hGM-CSF (produced in 293T cells) in RPMI with 10% heat-inactivated, pooled human AB serum (Sigma) for 7–8 days. CD14 + monocytes were also differentiated into monocyte-derived Macrophages (hMDMs) using hM-CSF (Peprotech, Human Recombinant M-CSF, #300-25) in RPMI with 10% pooled human AB serum (Sigma) for 5–6 days.
BLaER1 were obtained from Dr. Veit Hornung laboratory, Munich^{30 (link),52 (link)} and cultured in RPMI, 10% FCS, 1% Glutamine, 1% Pyruvate + 1% Pen/strep. For differentiation, 10⁵ cells in culture media was used in 96 well plate supplemented with 10 ng/ml of hrIL-3 (PeproTech, Cat#200-03), 10 ng/ml hr-CSF-1 (M-CSF) (PeproTech, Cat#300-25), and 100 nM β-Estradiol (Sigma-Aldrich, Cat#E8875) and incubated for 5–6 days. Before stimulation, differentiation media was changed to culture media.