Gene expression analysis of myeloid cells cultured in the presence of fibronectin and/or anti-ILT3 16C5

RNA sequencing of monocyte-derived dendritic cells, tolerogenic dendritic cells, unpolarized macrophages, and M1, M2a, and M2c macrophages differentiated in vitro on PBS- or fibronectin-coated wells, in the presence of control IgG1 or anti-ILT3 16C5 Overall design: Peripheral blood monocytes from healthy human donors were plated on wells pre-coated with PBS or fibronectin (5 ug/mL) in the presence of an anti-KLH control antibody or anti-ILT3 16C5 (10 ug/mL). Monocytes were differentiated into dendritic cells or macrophages as follows: Monocyte-derived DCs (GM-CSF/IL-4 for 7 days), tolerogenic DCs (GM-CSF/IL-4 for 5 days + vitamin D3/dexamethasone for 2 days), unpolarized macrophages (M-CSF for 7 days), or polarized macrophages (M-CSF for 5 days plus IFN-g/LPS for 2 days for M1 macrophages, IL-4 for 2 days for M2a macrophages, or IL-10 for 2 days for M2c macrophages). Cells were harvested in TRIzol at day 7, and RNA was purified using the Qiagen RNeasy kit. Total RNA was prepared for sequencing using the TailorMix Directional RNA Sample Preparation Kit (SeqMatic) and sequenced on an Illumina NovaSeq S1.