Figure 1
ROS production after CD40 ligation in Raji human B cells. (A) The cells (1 × 106) that were incubated with 20 µM DCFDA for 15 min were stimulated with either control Ig or anti-CD40 monoclonal Ab (10 µg/ml). When pretreated with the inhibitors, the cells were preincubated in the presence of inhibitors for 30 min before the addition of DCFDA. Representative microscopic fields of 2', 7'-dichlorofluorescein (DCF) fluorescence from the Raji cells after a 20-min stimulation with anti-CD40 in the absence or presence of treatment with several inhibitors (15 µM DPI, 35 µM ETYA, 0.5 µM MK-886, 10 µM LY294002, or 0.1 µM wortmannin). Fluorescence of the cells after stimulation with control Ig or CD40 ligation is shown. Data represent three independent experiments. (B) The DCF fluorescence intensity in Raji B cells after a 20-min stimulation with anti-CD40 is shown. The different treatments are indicated. The numbers of cells (n = 35) on the fluorescence image from all three experiments are given. The data are presented as mean ± SD. Statistical significance was determined by two-way analysis of variance, and the data was subjected to paired t-tests as post-hoc analysis. ***, P < 0.001 indicates significantly lower values for cells under CD40 ligation than values for cells either untreated or treated with DPI.
