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FLT3L-cultured DCs (1 × 106) were stimulated with either LPS (10 μg/ml; Sigma-Aldrich) or CpG (200 nM; Invivogen) for 2 hours, treated with GolgiStop (BD Bioscience), and then incubated for another 2 or 4 hours. Cells were washed before being analyzed for cell surface and intracellular antigens. For analysis of CD80, CD86, or CD40 expression, cells were incubated with CpG or LPS for 24 hours before surface staining.
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