Foxp3 alexa fluor 488 cd25 pe cd4 percp

Spleen cells were measured after 5 days of stimulation. The proportions of CD4+CD25+Foxp3+ (forkhead box P3) Tregs in the cell mixtures were analyzed by flow cytometry using a True-Nuclear One-Step Staining Mouse Treg Flow kit (FOXP3 Alexa Fluor 488/CD25 PE/CD4 PerCP; Biolegend). The results are expressed as the percentages of CD4+CD25+Foxp3+T cells in CD4+ cells.

Mouse BMCs were seeded in 24-well plates and then left untreated or treated with S. aureus biofilm. After 48-hr treatment, the CD11b-positive cell populations were isolated from the untreated or biofilm-treated BMCs. The isolated CD11b-positive cell populations were then cocultured with spleen T cells that were stimulated by Dynabeads Mouse T-Activator CD3/CD28 (Gibco). After 24- or 48-hr coculture, the proportions of Foxp3⁺CD25⁺CD4⁺ Tregs in the cell mixtures were analyzed by flow cytometry using a True-Nuclear One Step Staining Mouse Treg Flow kit (FOXP3 Alexa Fluor 488/CD25 PE/CD4 PerCP; Biolegend). Results were expressed as the percentages of Foxp3⁺CD25⁺CD4⁺ T cells in CD4⁺ cells.