Fig. 1

Flow cytometry gate-strategy to identify non-classical and classical monocyte subsets and myeloid dendritic cells (mDCs). In a the conventional gating strategy is shown, representing classical, intermediate and non-classical monocyte populations based on CD14 and CD16 expression. Since after LPS stimulation CD16 is downregulated, CD33 was used combined with CD14 to distinguish the classical and non-classical monocytes: R1 classical monocytes (CD14⁺⁺CD33⁺⁺) is equivalent to CD14⁺⁺CD16⁻; R2 non-classical monocytes (CD14^+/−CD33^+/dim) correspond to CD14⁺CD16⁺⁺ (b–c). mDCs (R3) were identified based on the following phenotype: CD14⁻CD33⁺⁺HLA-DR⁺⁺ (a and d). Monocytes and mDCs characteristics of forward scatter (FSC), side scatter (SSC) and CD45 expression and therefore lymphocytes populations are excluded from the analyses (e–f)