The expression levels of RelB in DCs under different conditioned microenvironments. Cells were lysed with RIPA buffer (20 mM sodium phosphate, pH 7.4, 150 mM sodium chloride, 1% Triton X-100, 5 mM EDTA, 200 μM phenymethylsulfonyl fluoride, 1 μg/ml aprotinin, 5 μg/ml leupeptin, 1 μg/ml pepstatin and 500 μM Na3VO4). The protein extracts were electrophoresed on 12% ~ 14% SDS-polyacrylamide gel and transferred onto a nitrocellulose membrane (Invitrogen, USA). After blocking with 5% BSA in 0.1% Tween 20 in PBS, membranes were probed with primary antibodies. Anti-RelB and anti-β-actin antibodies (Sigma) were diluted in blocking buffer and incubated with the blots overnight at 4°C. The bound primary antibodies were probed with a 1:2000 diluted secondary antibody (goat anti-human IgG-HRP antibody) and visualized by the ECL chemiluminescence system (Amersham, USA). The gray values of proteins were measured by Image J (1.45). The expression levels of proteins were normalized to those of corresponding β-actin. Compared with DCs: *
P < 0.05 or **
P < 0.01.