Alexa fluor 647anti mouse foxp3 catalog 126408

FITC anti-mouse CD29 (Catalog #102205), FITC anti-mouse Sca-1 (Catalog #108105), PE anti-mouse CD34 (Catalog #119307), PE anti-mouse CD45 (Catalog#103105), FITC anti-mouse CD4 (Catalog #100509), and Alexa Fluor 647anti-mouse FOXP3 (Catalog #126408) were purchased from BioLegend (San Diego, USA). FACS CantoII (BD Biosciences) was used for flow cytometry, and data were analyzed by BD FACSDiva software (BD Biosciences) and FlowJo (v X.0.7) software.

Splenocytes were freshly isolated from the spleens of recipient mice. Briefly, the spleen was mashed through a cell strainer and centrifuged at 1000 rpm for 5 min. Then, the cells were washed in cell staining buffer (BioLegend) and centrifuged at 1400 rpm for 5 min. The red blood cells were lysed by ammonium chloride solution (Stemcell) for 10 min, followed by washes and centrifugation. Finally, the cells were stained by FITC anti-mouse CD4 (Catalog #100509) according to the Cell Surface Immunofluorescence Staining Protocol (BioLegend), followed by staining with Alexa Fluor 647 anti-mouse FOXP3 (Catalog #126408) according to True-Nuclear™ Transcription Factor Staining Protocol (BioLegend). After that, the percentage of CD4⁺ Foxp3⁺Treg cells was evaluated by flow cytometry.