Apc cyanine7 anti mouse cd3 antibody

Flow cytometry was performed as previously described (Zhao et al., 2018 (link)). Briefly, kidneys were weighed and minced. A collagenase solution (1 mg/ml; Sigma-Aldrich, St. Louis, MO, United States) was used for renal digestion for 30 min at 37°C. A 100-μm cell strainer (Fisher Scientific) was used, together with a 1-ml syringe plunger, to acquire single cells. Cell pellets were washed and resuspended for further experiments. Anti-Mouse CD16/CD32 (553141; RRID:AB_394656, clone 2.4G2; BD Biosciences, San Jose, CA, United States) was used for non-specific Fc block. Antibodies used in this assay were acquired from BioLegend, San Diego, CA, United States; they include BV421 anti-mouse CD45 (RRID:AB_2562559), APC anti-mouse F4/80 antibody (RRID:AB_893481), PE anti-mouse CD206 (MMR) antibody (RRID:AB_10895754), FITC anti-mouse CD86 antibody (RRID:AB_313149), PerCP/Cyanine5.5 anti-mouse/human CD11b (RRID:AB_893232), APC/Cyanine7 anti-mouse CD3 antibody (RRID:AB_2242784), FITC anti-mouse CD4 antibody (RRID:AB_312713), and APC anti-mouse CD8a antibody (RRID:AB_312750). Data were acquired on a FACS Calibur cytometer [Becton Dickinson (BD), Bedford, MA, United States] and analyzed using FlowJo software (Tree Star, Ashland, OR, United States).