Effects of inhibitors on activation of signal molecules in co-cultured-U87 cells. Experimental details in co-culture were indicated in Figure 1. The anti-CD40 antibody (300 ng/ml), Jak inhibitor (10 μM AG490) or 8-oxo-dG (300 μg/ml) was pretreated in astrocytes 1 h, 5 and 10 min, respectively, before co-culture, and CD40 siRNA was transfected, as described in "Methods". Activities of PKCs, MAP kinases and transcription factors were determined in protein extracts and nuclear extracts by Western blot and EMSA, respectively. (A, B) Activities of PKC isoforms and MAP kinases after inhibitor pretreatment. (C, D) Activities of NF-κB (upper panel) or AP-1 (lower panel) and expression of CBP after inhibitor pretreatment. *, Numbers below bands are mean values obtained from four independent experiments (n = 4) as the ratio of band density of each group versus that of total protein or actin using densitometry analysis.