Figure 6.

IL-1β-induced CamKII activation and effect of KN-62 on IL-1β-induced IL-6 synthesis in primary cortical neurones. (a) Western blot analysis of CamKII activation in total cell extracts of cultures treated with vehicle (C) or IL-1β (0.3 U ml⁻¹) for 5 or 15 min, in the absence or presence of IL-1RA (50 ng ml⁻¹), 3OMS (25 μM), PP2 (10 μM), KN-62 (CamkII/IV inhibitor, 10 μM) or AP5 (50 μM). Data are presented as the mean±s.d. of five different experiments in separate cultures, ^**P<0.01, IL-1β vs control, ^##P<0.01, IL-1β+inhibitor vs IL-1β alone. (b) Cultures were treated with IL-1β (0.3 U ml⁻¹) for 7 h in the absence or presence of KN-62 (10 μM). Cell lysates and supernatants were assayed for IL-6 using a specific mouse ELISA. Data are presented as the mean±s.d. of six different experiments in separate cultures. ^**P<0.01 IL-1β vs control, ^###P<0.001, IL-1β+KN-62 vs IL-1β alone. IL, interleukin; 3OMS, 3-O-methyl-sphingomyelin.