Anti fas pe

Single cell suspensions were stained using combinations of anti-CD11c-PE (N418, BD Biosciences, San Jose, CA, USA), anti-B220-PerCP (RA3-6B2, BD Biosciences), anti-I-A^b/Ea_52-68-biotin (YAe, eBioscience, San Diego, CA, USA), anti-CD4-eFluor450 (RM4-5, eBioscience), anti-ICOS-alexa-fluor-488 (C398.4A, BioLegend, San Diego, CA, USA), anti-PD-1-PE-Cy7 (J43, eBiosciences), anti-SLAM-APC (TC15-12F12.2, BioLegend), biotinylated anti-CXCR5 (2G8, BD Biosciences), anti-B220-eFluor450 (RA3-6B2, eBioscience), anti-GL-7-FITC (BD Biosciences), anti-FAS-PE (BD Biosciences) and biotinylated anti-IgM^a (DM-1, BD Biosciences). Biotinylated antibodies were detected by incubation with fluorochrome-conjugated streptavidin (BD Biosciences). Appropriate isotype controls were used throughout. Samples were acquired using a MACSQuant analyzer (Miltenyi Biotec, Surrey, UK) and analysed using FlowJo software (Tree Star Inc, Ashland, OR, USA).

Antibodies were purchased as followed: Anti-CD4-V450, anti-CD8α-APCcy7, anti-B220-V450, anti-CD138-Pe-cy7, anti-CD86-Pe-cy5, anti-FAS-PE, anti-GL-7-APC, anti-CD40-APC, anti-MHCII-FITC, anti-IL-4-PE, anti-IL-5-PE, anti-IgM-Pe-cy7, anti-IgG1-APC, anti-IFN-r-Percp5.5, anti-TNF-α-PE, and anti-IL-17-Pe-cy7 were purchased from BD Biosciences (Franklin Lakes, NJ). Anti-PDL-1-biotin (eBioscience, San Diego, CA), anti-XBP-1s (Cell Signaling Technology, Danvers, MA), and anti-Rabbit IgG-FITC (Thermo Fisher Scientific, Waltham, MA) antibodies were purchased from commercial sources. Recombinant mouse IL-4 (PeproTech, Rocky Hill, NJ) and LPS (Sigma-Aldrich, St. Louis, MO) were purchased from the commercial companies. Goat F(ab’)2 Anti-Mouse IgM (1022-01, SouthernBiotech, Birmingham, AL) and anti-mouse CD40 (BE0016-2, BioXCell, Lebanon, NH) were commercially purchased from companies.