Fig 1: C5a affects the pro-inflammatory potential of 6-sulfo LacNAc dendritic cells (slanDCs). (A) Slan, CD123, CD1c, CD141, CD11c, HLA-DR, and CD14 expressions in the 120 different clusters defined by spanning-tree progression analysis of density-normalized events (SPADE). (B) C5aR expression in the 120 clusters defined by SPADE. Different dendritic cell (DC) subsets are indicated with black ellipses and were selected based on the expression of known DC identification markers (12, 13). pDCs were selected as CD123+, CD11c−, CD1c−, CD14−, and slan−; slanDCs were CD11c+, CD1c−, CD14−, slan+, and CD123−; MDC1s were CD11c+, CD1c+, CD14−, slan−, and CD123−; and MDC2s were CD11c+, CD141++, CD1c−, CD14−, and slan−. (A,B) Fluorescent intensity of marker expression is visualized using a color scale. (C) Sorted slanDCs were stimulated overnight with LPS or R848 or left untreated. The production of TNF-α and IL-12p40 in supernatants is depicted (n = 4). (D) The production of TNF-α and IL-12p40 by slanDC determined after overnight stimulation with R848 in the absence or presence of C5a (n = 5). Cytokine production was measured using ELISA.