Transcriptomic profile of monocytes/ macrophages from injured bone, muscle, skin or heart tissues of Treg-depleted mice

Mice were depleted of Tregs, and bone, muscle, skin or heart injuries were performed. On day 4 and 7 post-injury, the tissues were harvested and the endogenous monocytes/macrophages were sorted using FACS. These were then used for bulk RNA sequencing, to compare the transcriptomic profiles of injured tissue monocytes/ macrophages in Treg-depleted mice versus control mice post-injury. Ten-week-old Foxp3(DTR/GFP) (FDG) mice and wild-type (WT) C57BL/6J (control) mice received intraperitoneal injections of 20 ng/g diphtheria toxin (DT), starting two days prior to the induction of injury, and continuing daily until the day of injury, followed by every 2 days after injury, for 1 week. Bone (cranial defect), muscle (volumetric muscle loss defect), skin (full-thickness biopsy punch) or heart (myocardial infarction) injuries were induced in 10 week old Foxp3(DTR/GFP) (FDG) mice and control C57BL6/J wild-type mice, that were treated with DT to deplete Tregs in the FDG mice. Only 1 injury per mouse was performed. On day 4 and 7 post-injury, endogenous monocytes/ macrophages were sorted from Foxp3(DTR/GFP) + DT mice (i.e FDG+DT, Treg-depleted), and wildtype C57BL/6J + DT mice (i.e. WT+DT, control), and used for bulk RNA sequencing.