Figure 3

PKCδ is involved in thrombin-induced HO-1 expression in synovial fibroblasts. (A, C) Osteoarthritis synovial fibroblasts (OASFs) (six OA lines) were pretreated for 30 minutes with GF109203X (3 μM), rottlerin (10 μM), and Ro320432 (10 μM) followed by stimulation with thrombin for 24 hours, and HO-1 expression was examined with Western blotting and qPCR. (B) Cells (seven OA lines) were transfected for 24 hours with PKCδ siRNA followed by stimulation with thrombin for 24 hours, and HO-1 expression was examined with qPCR. (C) Cells (six OA lines) were incubated with thrombin for indicated time intervals, and PKCδ phosphorylation was examined with Western blotting. (D, E) Cells (seven OA lines) were incubated with thrombin for 30 minutes or pretreated for 30 minutes with PPACK or transfected 24 hours with PAR1 and PAR3 siRNA, followed by stimulation with thrombin for 30 minutes, and PKCδ kinase activity was determined with the PKCδ kinase kit. Results are expressed as the mean ± SEM. *P < 0.05 as compared with basal level. #P < 0.05 as compared with thrombin-treated group.