Fitc anti human cd68

Single-cell suspensions of TNMs and TAMs were prepared as described above and stained with BUV395 anti-human-CD45, APC anti-human CD11b, FITC anti-human-CD68, or BV-480 anti-human-CD206 (all antibodies were purchased from BD Biosciences). Then, labeled cells were analyzed on a FACSCanto II Flow Cytometer (BD Biosciences), and the data were processed using FlowJo Version 10.0. For the phenotypic analysis of THP-1-derived M2-polarized macrophages, cells were stained with APC anti-human-CD11b and FITC anti-human-CD206.
In addition, immunofluorescent staining was performed on TNMs and TAMs using anti-F4/80 and anti-CD206 antibodies (all purchased from Abcam, UK).

Hypoxia green reagent; pHrodo Green E. coli; DAF-FM DA; JSH-23; QNZ; ABT-263; and ABT-737 were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The fetal bovine serum was from Gibco (Gibco, Sigma Aldrich Company Ltd., Waltham, MA, USA). Intracellular staining permeabilization wash buffer; Human TruStain FcX (Fc Receptor Blocking Solution); PE anti-human Ki-67; isotype control antibodies PE Mouse IgG1 k isotype Ctrl; APC Mouse IgG1 k isotype Ctrl; FITC Mouse IgG1 k isotype Ctrl; PE Mouse IgG2a k isotype; PE Mouse IgG2b k isotype and antibodies APC anti-human CD11b; PE anti-human CD284; PE anti-human CD36; PE anti-human CD33; PE anti-human HLA-DR; PE anti-human HIF-1α were from BioLegend (San Diego, CA, USA). MycoFluor™ Mycoplasma Detection Kit was from Molecular Probes Inc. (Eugene, Oregon, USA). FITC anti-human CD68 was from BD Bioscience (Franklin Lakes, NJ, USA). Culture medium RPMI 1640; 2-mercaptoethanol; PBS; Calcein AM; Bisbenzimide Hoechst 33342 (H33342); propidium iodide (PI), resazurin; LPS of E. coli O111: B4; antibodies FITC anti-human CD11c; FITC anti-human CD14; FITC anti-human CD54; NF-kB activation inhibitor IV, and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).