Preparation of nuclear extract and colorimetric NF-κB assay.

DC were stimulated with different doses of anti-Tim-1 or rIgG2a, or LPS (200 ng/ml) plus CpG (500 nM), and nuclear extracts were prepared using a nuclear extract kit (Active Motif, Carlsbad, CA). NF-κB/DNA binding activity was detected using a TransAM NF-κB p65 transcription factor assay kit (Active Motif) according to the manufacturer’s protocol.