For fluorescence imaging of frozen sections, mice were given the same treatments as those in the DC maturation study, and lymph node sections were prepared as described above. Then, the sections were labeled with an anti-Ki67 antibody followed by a fluorophore-labeled secondary antibody and then analyzed with an automatic multispectral imaging system (PerkinElmer).

For CFSE-based FCM analysis, mice were given the same treatments as those in the DC maturation study, and then, T cells from the spleens and lymph nodes of the mice in each group were labeled with CFSE. Then, CFSE-labeled T cells were intravenously injected into the mice that had been treated in the same way as the mice from which T cells were derived. Three days later, the inguinal lymph nodes and spleen of each mouse were collected and triturated into single-cell suspensions. Then, after labeling with an APC/Cy7-conjugated anti-mouse CD3 antibody and a PE-conjugated anti-mouse CD8a antibody and washing, the cells were dispersed in 500 μl of PBS and analyzed by FCM.

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