Figure 3

PPCSE-induced HO-1 expression is mediated by PKCδ in bEnd.3 cells. A-D: Cells were pretreated with non-selective inhibitors of PKC, GF109203X (GFX) and Ro318220 (Ro), and Gö6976 (Gö) or selective inhibitor of PKCδ, rottlerin (Rott) for 1 h, and then incubated with PPCSE for 24 h. The expression of HO-1 was determined by Western blot. E: Cells were transfected with a dominant negative mutant of PKCδ (ΔPKCδ), and then incubated with PPCSE for 24 h. The expression of HO-1 was determined by Western blot. F: Cells were treated with 30 μg/ml PPCSE for the indicated time intervals, and then the activity of PKC was determined as described in the Methods. G: Cells were pretreated with rottlerin (Rott) for 1 h, and then incubated with PPCSE for 1 h. The ROS generation and NADPH oxidase activity were measured. Data are expressed as the mean ± SEM of three independent experiments performed in triplicate. A-F: ^# P < 0.05 as compared with the cells exposed to PPCSE alone. G: ^# P < 0.05 as compared within groups.