Clone t45 2342

BALB/c mice were infected with 1,000 tachyzoites of WT, KO P18 or CPT P18 (48 (link)) and sacrificed on day 4 post-tachyzoites injection. Peritoneal lavage was performed. Cells recovered from the peritoneum and spleen-derived murine cells from BALB/c mice infected with either parasite line, were stained for 20 min in dark, with anti-CD11b⁺ directly conjugated to Phycoerythrin-Cy7 (PE-Cy7) (BD Bioscience, 2 μg/ml, Clone M1/70), anti-CD11c+ directly conjugated to PE (BD Biosciences, 2 μg/ml, Clone HL3), F4/80 directly conjugated to PE (BD Biosciences, 2 μg/ml, Clone T45-2342) or CD335+ (NKp-46) (BD Biosciences, 2 μg/ml, clone 29A1.4), directly conjugated to Phycoerythrin (PE). Labeled samples were washed twice and 10,000 events were acquired using a FACS ARIAII cell sorter. The flow cytometry results reflect the percentage of different populations of cells in the peritoneum or the spleen (as indicated). Debris and dead cells are gated out by Forward Scatter (FSC) versus Side Scatter (SSC) gating.

Cells were counted (5 × 10⁶ cells/per sample) and stained for 20 min at 4℃ Fixable viability stain 510 (Cat. #564406, 2 mg/mL, BD Biosciences) for live cell staining, then the surface molecules of the cells were stained with antibodies for 30 min at 4℃. The panel of antibodies designed to analyze of myeloid cells: Alexa Fluor 700-CD45 (Cat. #560510, Clone 30-F11, 1 mg/mL, BD Biosciences), BV421-F4/80 (Cat. #565411, Clone T45-2342, 1 mg/mL, BD Biosciences), BV605-CD11b (Cat. #557672, Clone M1/70, 1 mg/mL, BD Biosciences), PE-MHC-II (Cat. #557000, Clone M5/114, 1 mg/mL, BD Biosciences), PE-CY7-CD11c (Cat. #117317, Clone N418, 1 mg/mL, BioLegend) and APC-CD206 (Cat. #17-2061-82, Clone MR6F3, 2 mg/mL, eBioscience). Samples were analyzed using Flow Cytometer (BD Biosciences). Subsequent analysis was performed with FlowJo software (Tree Star Inc.).