Figure 4.

Impaired phosphorylation of p100 after CD40 ligation in mature DCs from NIK-mutated aly/aly mice. A primary culture of DCs, SDDCs were prepared from splenocytes from aly/+ or aly/aly mice. Unstimulated or LPS-treated SDDCs were considered as immature (iDC) or mature DCs (mDC), respectively. (a) Cell surface marker expressions. The iDC and mDC from aly/+ or aly/aly mice were stained with specific mAb against CD40, CD86, or MHC class II (I-A^b) or isotype-matched control immunoglobulin (Isotype). The cell surface expressions were analyzed by flow cytometry. A representative histogram of three independent experiments is shown (upper). The mean fluorescence intensity (MFI) of the cells is represented (lower). Each column represents the mean ± SE of three independent experiments. (b) Phosphorylation of p100, p38 MAPK. The mature DCs from aly/+ or aly/aly mice were treated with anti-CD40 mAb (α-CD40) or control immunoglobulin for 20 min, and then whole cell lysates were prepared. Levels of phospho-p100 (pp100), p100, phospho-p38 MAPK (pp38), and p38 MAPK (p38) in the cell lysates were determined by immunoblotting. A representative immunoblot of three independent experiments is shown (upper). The relative intensity of the specific band is represented (lower). Each column represents the mean ± SE of three independent experiments.