Innate Immune Training of Osteoclastogenesis promotes Inflammatory Bone Loss

scRNAseq of myeloid cells from arthritic hind paws upon induction of trained immunity or control treatment CD45+CD11b+ synovial myeloid cells from hind paws of b-glucan or PBS pre-treated mice (7 days prior to K/BxN STA induction) were sorted at day 17 after arthritis induction using a FACS Aria III sorter (BD, Heidelberg, Germany). The cell concentration was adjusted to 1000–1600 cells per μl (1.537 cells/μl in PBS sample and 1150 cells/μl in b-glucan sample). Approximately 10.000 cells were loaded and then processed with Chromium Next GEM Single Cell 3' v3.1 dual-index kit (10X Genomics) at the Dresden Concept Genome Center, Dresden, Germany. According to the manufacturer’s instructions, the cDNA was synthesized and subjected to library preparation and sequencing on the Novaseq 6000 platform (Illumina).