Figure 5.

RelB down-regulation leads to diminished APC function. (A) EMSA and supershift analysis of nuclear extracts prepared from DG75 incubated for 24 h in the presence or absence of 50 ng/ml sCD40L. (B) DG75 lines stably transfected with pAntiRelB or pCDNA3 were incubated with or without sCD40L for 0 or 4 h, then cytospun. Slides were fixed in paraformaldehyde and stained with anti-RelB by using diaminobenzidine as the chromogen (black, magnification ×40). (C) pAntiRelB or pCDNA3 DG75 lines were incubated in the presence (bold line) or absence (thin line) of 50 ng/ml sCD40L, then stained for expression of MHC class I and MHC class II. Isotype controls are indicated by the filled histograms. Significant differences were observed in levels of MHC class I (see Supplementary Fig. 7C at www.pnas.org) (P = 0.0202) and MHC class II (see Supplementary Fig. 7D at www.pnas.org) (P = 0.0463) after CD40L treatment of pCDNA3-DG75 but not for pAntiRelB-DG75. Data are representative of three separate experiments. (D) Stably transfected pAntiRelB- or pCDNA3-transfected DG75 were incubated in the presence or absence of 50 ng/ml sCD40L for 24 h, washed, and used in MLR as described. Data are the mean of triplicate wells ± SD and are representative of three separate experiments.