The C5a signaling triggers functional intracellular signaling in C5aR+ LysoDCs

C5a–C5aR signaling results in an intracellular calcium influx through activation of the coupled G-protein, which triggers ROS generation in APCs. To investigate the effect of C5a–C5aR signaling on cell function, we analyzed the gene expression profile of rC5a-treated C5aR+ LysoDCs. PP cells were prepared by collagenase D (Roche)/DNase I buffer and after removing dead cells using a dead cell removal kit, CD11c+ cells were enriched using anti-mouse CD11c microbeads. Sorted C5aR+ LysoDCs were stimulated with recombinant C5a protein (100 nM) for 3 hours. Total RNA was prepared using an RNeasy PLUS Micro kit according to the manufacturer’s protocols (Qiagen).