Apc conjugated anti mouse f4 80

Cell surface staining was conducted with the appropriate fluorochrome-conjugated Abs for 30 min at 4°C. The following specific monoclonal antibodies (mAbs) were used: antigen-presenting cells (APC)-cy7-conjugated anti-mouse CD45, FITC-conjugated anti-mouse CD3, Percy5.5-conjugated anti-mouse CD4, FITC-conjugated anti-mouse CD8, PEcy7-conjugated anti-mouse NK1.1, BV421-conjugated anti-mouse Nkp46, BV421-conjugated anti-mouse major histocompatibility complex II (MHC-II), APC-conjugated anti-mouse F4/80, PE-conjugated anti-mouse CD11c, PE-CF594-conjugated anti-mouse CD11b and APC-conjugated anti-mouse CD27 (all purchased from BD Bioscience). Flow cytometric (FCM) analyses were performed on a CyAn ADP analyzer (Beckman Coulter, Inc., Kraemer Boulevard Brea, CA, USA), with data analyzed using FlowJo Version 6.1 software (TreeStar, Asland, OR, USA).

Isolated pulmonary macrophages were incubated with FITC‐conjugated anti‐mouse CD80 (BD Biosciences, CA, USA), APC‐conjugated anti‐mouse F4/80 (BD Biosciences), and PerCP‐Cy5.5‐conjugated CD206 (BD Biosciences). Analysis was performed by ImageStreamx Mark II flow cytometer (Merck, Darmstadt, Germany). M1 macrophages were classified as F4/80+/CD80+ and M2 macrophages were identified as F4/80+/CD206.