Pe cy7 anti mouse cd25 antibody

1,3-propanediol, p-toluenesulfonic acid, ethyl acetate, and sodium sulfate (Na2SO4), as well as dicyclohexylcarbodiimide (DCC), n-hydroxysuccinimide (NHS), and dimethylformamide (DMF) were purchased from Beijing Qiangsheng Technology Co., Ltd. 1MT was obtained from Sigma-Aldrich. GA and cell proliferation EdU image kits were purchased from Dalian Meilun Biotechnology Co., Ltd. The Annexin V-FITC/PI cell apoptosis detection kit was purchased from KeyGEN BioTECH and the Cell Counting Kit-8 (CCK-8) was from Beyotime Biotechnology. The radio immunoprecipitation assay lysis buffer, bicinchoninic acid protein assay kit, tyrosinase, proliferating cell nuclear antigen (PCNA) rabbit monoclonal antibody, cleaved caspase-3 rabbit monoclonal antibody, and horseradish peroxidase (HRP)–conjugated affinity Goat anti-rabbit/mouse immunoglobulin G (IgG) (H+L) were purchased from Beyotime Biotechnology. Matrigel was purchased from BD Biosciences, USA. Fluorescein isothiocyanate anti-mouse CD3 antibody, phycoerythrin (PE) anti-mouse CD8 antibody, antigen-presenting cell (APC) anti-mouse CD4 antibody, PE/Cy7 anti-mouse CD25 antibody, PE anti-mouse Forkhead box protein P3 (FOXP3) antibody, and APC anti-mouse natural killer (NK) 1.1 antibody were obtained from BioLegend Inc.

AT-infiltrating stromal cells were stained with FITC anti-mouse CD3ε antibody, PE anti-mouse CD4 antibody, PE/Cy7 anti-mouse CD25 antibody, APC anti-mouse CD8a antibody, FITC anti-mouse CD127 antibody, PerCP/Cy5.5 anti-mouse CD4 antibody, APC anti-mouse CD25 antibody (these antibodies were purchased from Biolegend, San Diego, CA, USA), PE anti-mouse/Foxp3 antibody (Invitrogen, Carlsbad, CA, USA), PE anti-mouse CD11b antibody (Biolegend), PE/Cy7 anti-mouse Ly-6C antibody (Biolegend, San Diego, CA, USA) and APC anti-toll-like receptor 4 (TLR4) antibody (Invitrogen, Waltham, MA, USA), and analyzed using an Accuri C6 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA).