Figure 1.

CD40 receptor activation by mCD40L but not sCD40L induces NORE1A expression in carcinomas. (a) The CD40-positive bladder carcinoma cell lines EJ, 253J and the CD40-negative MGHU3 bladder carcinoma cells and the CD40-positive pancreatic cell line Panc1 and the CD40-negative MiaPaca-2 cells were infected with 100 and 30 MOI of either RAdMock (AdM) or RAdnCD40L (AdnL), respectively, or left uninfected as a negative control or treated with sCD40L (sL) at a final concentration of 1 μg/ml for 24 h. Cells were then examined for CD40 expression by FACS analysis and mCD40L, NORE1A expression by western blot analysis. β-Actin expression was examined to ensure equal loading between different samples. (b) EJ cells were infected with 100 MOI of either RAdMock or RAdnCD40L or left untreated as a negative control or treated with sCD40L at a final concentration of 1 μg/ml for the indicated time points. Cells were lysed and examined for the expression of mCD40L, NORE1A, IκBα and the β-actin as a loading control. The phosphorylated levels of JNK, AKT, p38 and ERK were also examined. (c) EJ cells were infected with 100 MOI of either RAdMock or RAdnCD40L or left untreated as a negative control or treated with sCD40L (1 μg/ml) for the indicated time points. Total cell lysates were examined for the expression of p-p65 and p-IκBα, IκBα and the β-actin as a loading control