Pe conjugated anti mouse cd11b

Concanavalin A (ConA), dexamethasone, and DPI were purchased from Sigma-Aldrich (St. Louis, MO). Murine IL-6 and GM-CSF cytokines were from PEPROTECH (Rocky Hill, NJ). PerCP/Cy5.5-conjugated anti-mouse (Gr-1), APC-conjugated anti-mouse CD3, and PE-conjugated anti-mouse CD11b were purchased from Biolegend (San Diego, CA). FITC-conjugated rat anti-mouse CD4, APC-conjugated anti-mouse CD69, and mouse regulatory T cell Staining Kit were purchased from eBioscience (San Diego, CA). Purified NA/LE rat anti-mouse CD25 was purchased from BD Pharmingen. Collagenase Type II and DNase I were purchased from Gibco by life technologies (Carlsbad, CA). Rabbit polyclonal antibody to Arginase-1 was purchased from Cell Signaling Technology (Danvers, MA) and mouse monoclonal antibody to alpha tubulin was purchased from Abcam (Cambridge, MA).

4-EG, triphenyltetrazolium chloride (TTC), Alexa Fluor 594 goat-anti rabbit IgG secondary antibody, Evans blue, Ficoll PM 400, and LPS (Escherichia coli O55:B5) were purchased from MilliporeSigma (St. Louis, MO, USA). Zinc protoporphyrin (ZnPP) was purchased from Alfa Aesar (Haverhill, MA, USA). 7-aminoactinomycin D (7-AAD), Alexa Fluor 488-conjugated anti-mouse CD45 (clone: 30-F11), APC-conjugated anti-mouse CD45 (clone: 30-F11), PE/Cy7-conjugated anti-mouse CD11b (clone: M1/70), PE-conjugated anti-mouse CD11b (clone: M1/70), PE/Cy7-conjugated anti-mouse CD86 (clone: GL-1), PE/Cy7-conjugated anti-mouse CD68 (clone: FA-11), Alexa Fluor 647-conjugated anti-mouse CD31 (clone: MEC13.3), PE/Cy7-conjugated anti-mouse ICAM-1 (clone: HA58), and PE/Cy7-conjugated anti-mouse VCAM-1 (clone: 429, MVCAM.A) antibodies for flow cytometer analysis were purchased from BioLegend (San Diego, CA, USA). PE-conjugated anti-mouse E-selectin (clone: 10E9.6) antibody for flow cytometry analysis was purchased from BD Biosciences (San Diego, CA, USA). Alexa Fluor 488 anti-mouse Iba1 (EPR16588) antibody for immunohistochemistry (IHC) was purchased from Abcam (Cambridge, MA, USA). Anti-mouse HO-1 (10701-1-AP) antibody for flow cytometer and IHC was purchased from Proteintech (Chicago, IL, USA).

Total cell number of BALF was counted, and total protein concentration in BALF was determined using a BCA assay Kit (Thermo Fisher Scientific) according to the manufacturer's instructions. The inflammatory cells in BALF were analyzed with a Cytoflex machine (Beckman Coulter), and the following fluorescence-conjugated antibodies were used for the experiment: PE-conjugated anti-mouse CD11b (BioLegend, San Diego, CA, USA), FITC-conjugated anti-mouse F4/80 (BioLegend, San Diego, CA, USA), and BV650-conjugated anti-mouse LY-6G (Invitrogen, Carlsbad, CA, USA).