Fig. 4.

a NF-κB activation in Mo-DCs-Tum. Six hours after GCTM-1 cell pulsation, nuclear translocation of NF-κB in Mo-DCs was detected by EMSA. Lane 1 Mo-DCs (control), Lane 2 Mo-DCs-Tum-A, Lane 3 Mo-DCs-Tum-N, Lane 4 competition assay by the addition of 100 times NF-κB oligonucleotide. Data are the ratio of the intensity of Mo-DCs-Tum to control Mo-DCs as determined by NIH image. b NF-κB activation in Mo-DCs-Tum in the presence of anti-TNF-α neutralizing antibody (1 μg/ml). Twelve hours after GCTM-1 cell pulsation, nuclear translocation of NF-κB p65 in Mo-DCs-Tum was analyzed by EMSA. Lane 1 Mo-DCs-Tum-A, Lane 2 Mo-DCs-Tum-A in the presence of anti-TNF-α neutralizing antibody, Lane 3 Mo-DCs-Tum-N, Lane 4 Mo-DCs-Tum-N in the presence of anti-TNF-α neutralizing antibody. Data are the ratio of Mo-DCs-Tum with anti-TNF-α neutralizing antibody to Mo-DCs-Tum without anti-TNF-α neutralizing antibody as determined by NIH image. c NF-κB activation in Mo-DCs-Tum in the presence of anti-IL-12 neutralizing antibody (20 μg/ml). Twelve hours after GCTM-1 cell pulsation, nuclear translocation of NF-κB p65 in Mo-DCs-Tum was analyzed by EMSA. Lane 1 Mo-DCs-Tum-A, Lane 2 Mo-DCs-Tum-A in the presence of anti-IL-12 neutralizing antibody, Lane 3 Mo-DCs-Tum-N, Lane 4 Mo-DCs-Tum-N in the presence of anti-IL-12 neutralizing antibody. Data are the ratio of Mo-DCs-Tum with anti-IL-12 neutralizing antibody to Mo-DCs-Tum without anti-IL-12 neutralizing antibody as determined by NIH image. The data are representative of three independent experiments using Mo-DCs generated from three different donors