Fitc conjugated anti cd11b m

Mononuclear cells were incubated with or without 50 ng/ml phorbol
myristate acetate (PMA) (Sigma) and 500 ng/ml ionomycin (Sigma) in the presence
of GolgiStop (BD) in complete T cell media at 37°C for 5 h.
Intracellular cytokine staining was performed according to the
manufacturer’s protocol. Cells were stained with Pacific Blue-conjugated
anti-CD4 (RM-5), PerCP-Cy5.5-conjugated anti-CD8a (53-6.7), APC-Cy7-conjugated
anti-TCRβ (H57-597), FITC-conjugated anti-CD62L (MEL-14), APC-conjugated
anti-CD44 (IM7), PE-conjugated-CD25 (PC61.5), PerCP-Cy5.5-conjugated-CD19
(eBio1D3), APC-conjugated anti-CD45.1 (A20), FITC-conjugated anti-CD45.2 (104),
Pacific Blue-conjugated anti-CD11c (N418), FITC conjugated-anti-CD11b (M1/70),
PerCP-Cy5.5-conjugated-anti-CD103 (2E7) (eBioscience), Biotin-conjugated
Vβ14 (14-2) (BD phamigen) and PE-Cy7-conjugated-Streptavidin (Thermo
Fisher Scientific). Cells were further stained intracellularly with
APC-conjugated anti-RORγ (B2D) (eBioscience) and PE-Cy7-conjugated
anti-IL-17a (eBio17B7) (eBioscience) using Foxp3 staining/permeabilization
buffer (eBioscience). Flow cytometric analysis was performed on an LSRII (BD
Biosciences). All data were re-analyzed using FlowJo (Tree Star).