2.3 Flow cytometry
To analyze the proliferation of T cells, CFSE (eBioscience Inc., San
Diego, CA, USA) was used. Cell surface MHC class II and CD86 on BMDCs
were stained with anti-I-A/I-E-PerCP (clone M5/114.15.2, BioLegend) and
anti-CD86-PE (clone GL-1, BioLegend), respectively. Intracellular IFN-γ
and IL-4 were stained with anti-IFN-γ-PE/Cyanine7 (clone XMG1.2,
BioLegend), and anti-IL-4-PE (clone 11B11, BioLegend), respectively,
with anti-CD4-FITC (clone GK1.5, BioLegend) after treatment with the
Fixation Buffer (#420801, BioLegend) and the Intracellular Staining
Perm Wash Buffer (#421002, BioLegend). Fluorescence was detected by a
MACS Quant Analyzer (Miltenyi Biotech) and analyzed with FlowJo (Tomy
Digital Biology, Tokyo, Japan).