Effect of isolated neutrophils from Cd177 knockout mice post-SCI on macrophages in a co-culture system

To investigate the transcriptional effects of CD177+ neutrophils on macrophages in spinal cord injuries (SCI), we adapted a co-culture system containing isolated neutrophils from Cd177 WT or KO mice post-SCI and bone marrow-derived macrophages (BMDMs). Each group had a biological repeat (n=3). Briefly, we isolated neutrophils in injured lesions at 3 dpi from Cd177 KO or WT mice by magnetic-activated cell sorting (MACS) and co-cultured them with BMDMs. After co-culturing for 24h, We isolated macrophages by fluorescence-activated cell sorting (FACS, gated on Ly6G-) for further testing.