Anti mouse cd80 pe clone 16 10a1

The following antibodies were used: anti-human CD11c-FITC (clone 3.9), anti-human CD86-PE (clone IT2.2), anti-human CD80-PE-Cy5 (clone 2D10.4), anti-human HLA-DR-PE-Cy5/Percp-Cy5.5 (clone LN3), anti-human CD123-PE (clone 6H6), anti-human CD274-PE (clone MIH1), anti-mouse CD11c-FITC (clone N418), anti-mouse MHC-II-PC5 (clone M5/114.15.2), anti-mouse 33D1-PE (clone 33D1), anti-mouse B220-PE (clone RA3-6B2), and anti-mouse CD274-PE (clone MIH5) were purchased from Invitrogen; anti-mouse CD86-PC5 (clone GL1) and anti-mouse CD80-PE (clone 16-10A1) were purchased from BD Biosciences. Prepared PBMCs or mice maternal-fetal interface mononuclear cells were washed with cold PBS containing 2% FBS and incubated with diluted antibodies at 4°C for 15 min. The cells were then washed twice with cold PBS containing 2% FBS, resuspended in 0.3 ml of PBS containing 2% FBS, and analyzed with a FACSVerse flow cytometer using the FACSuite software.

PD‐1 expression was detected using anti‐mouse PD‐1 APC (clone 29F.1A12; BioLegend, San Diego, CA, USA) and CD80 was detected using anti‐mouse CD80 PE (clone 16‐10.A1; BD Biosciences). Cells were processed and stained in fluorescence‐activated cell sorting (FACS) buffer comprising PBS supplemented with 0.1% bovine serum albumin (Sigma) and 0.1% sodium azide (Sigma). Cells were stained between 25 and 45 min on ice in the dark. Stained cells were analyzed on the FACS Fortessa X20 (Becton Dickinson).