Anti cd49b apc

On the third day of infection, peritoneal cells from WT mice infected with 1 × 10⁷ Nc-1 tachyzoites were collected and stained for phenotypic characterization and Dectin-1 expression. Briefly, mice were euthanized, and their peritoneal cavities were washed with ice-cold PBS. The suspension was then centrifuged at 400 × g, at 4°C, for 10 min. The cell pellet was resuspended in PBS with 5% of normal rabbit serum, at room temperature, for 15 min, prior to incubation with the appropriate antibodies: anti-CD11b-APC-Cy7, anti-CD11c-FITC, anti-MHCII-PE, anti-CD11c-V450, anti-CD19-APC-Cy7, anti-CD3-Pacific Blue, anti-CD49b-APC (BD Biosciences), and anti-Dectin-1-PE (R&D Systems, Minneapolis, MN, USA). Cells were incubated with primary antibodies conjugated to the different fluorochromes for another 30 min, at room temperature. After washing, cells were suspended in PBS with 3% formaldehyde, read by flow cytometry (FACSCantoII, BD Biosciences), and analyzed using dedicated software (FlowJo X, Tree Star Inc., Ashland, OR, USA).

Spleen and tumor cells obtained from control or treated tumor-bearing mice were thawed, centrifuged, and incubated with monoclonal antibodies conjugated with fluorophores: anti-CD45 V500, anti-CD11b PerCP-Cy5.5, anti-CD11c BV605, anti-CD4 APC, anti-B220 APC, anti-CD49b APC, anti-Ly6C PE, anti-Ly6G APC-Cy7, anti-MHCII FITC, and anti-CD86 PE-Cy7 (all from BD Biosciences). After incubation, cells were suspended in PBS with DAPI dye (Molecular Probes) and analyzed using LSR Fortessa with Diva Software (Becton Dickinson) according to the procedure described by Rossowska and coworkers (27 (link)).