Gl7 apc

The following Abs were used to stain splenocytes for flow cytometry: B220-BrilliantViolet(BV)605 (BioLegend), Ly5.2-AlexaFluor(AF)700, Ly5.1-PE-Cy7, GL7-APC, CD95-biotin, CCR6-biotin, CD38-FITC, CXCR4-PerCP-eFluor710, CD80-FITC, PD-L2-APC, and Y-Ae-biotin (all from eBioscience). Biotinylated Abs were detected with streptavidin-coupled BV421 (BioLegend). Additional reagents used for flow cytometry included PNA-FITC and NP-PE. Surface staining was carried out as described [29 (link)]. MitoTracker Red (Invitrogen), Annexin V-FITC (eBioscience) and SYTOX Red viability dye (Invitrogen) were used according to manufacturer’s instructions. Data were acquired on a LSRII flow cytometer (BD Biosciences) and analyzed using FlowJo software (Treestar, Inc.).

NS398, the inhibitor of COX-2 activity, was purchased from MedChemExpress (HY-13913, NJ, USA). Fluorescein-conjugated anti-mouse antibodies (F4/80-FITC, CD11b-PE, Gr-1-APC, Dead-APC-A750, CD45-PB450, CD45-KO525; CD4-FITC, CD3-PC5.5, IL-17A-PC7, IFN-γ-APC, IL-4-PB450; CD3-FITC, Foxp3-PC5.5, PD-1-PC7, CXCR5-APC, FVD-PB450, CD4-KO525; FAS-PE, CD19-PC5.5, B220-PC7, GL7-APC, CD138-APC-A750, FVD-PB450) and their corresponding isotype controls were obtained from eBioscience (San Diego, CA, USA). Ly6G-PC7 and Ly6G-allophycocyanin were obtained from BD Biosciences (San Jose, CA, USA).