Fig. 3. Effects of inhibitors of PKC and Raf-1 on phosphorylation of ERK and expression of CCL11 induced by thrombin. (A) HAo-SMCs were stimulated with thrombin for 5 min after incubation with or without GF109203X (3 µM) and GW5074 (25 nM) for 2 h. An equal amount of protein was subjected to Western blot analysis with antibodies against α-tubulin and phosphorylated and unphosphorylated forms of ERK1/2. (B, C) HAoSMCs were incubated with or without GF109203X (3 µM) and GW5074 (25 nM) for 2 h followed by stimulation with thrombin for 9 h. The levels of CCL11 transcripts were determined by realtime PCR (B), and the levels of CCL11 protein secreted into culture media were determined by ELISA (C). Data are expressed as mean±SD (n=3 replicates for each group). ^***p<0.001 vs. control; ^###p<0.001 vs. thrombin.