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. 2006 Apr 3;148(3):350–365. doi: 10.1038/sj.bjp.0706727

Figure 8.

Figure 8

PKCɛ but not PKCδ is involved in hGHSR-1a-mediated activation of ERK1/2. (a) CHO cells were transiently transfected with plasmid encoding hGHSR-1a (10 μg) together with 20 μg of empty pEGFP-N1 vector, wild-type PKCɛ-GFP or c-myc-tagged DN PKCɛ. Cells, which had been serum-starved for 1 h were stimulated with ghrelin (100 nM, 5 min). (b) CHO cells were transiently transfected with plasmid DNA encoding hGHSR-1a (10 μg) together with 20 μg of empty pEGFP-N1 vector, wild-type PKCδ-GFP or c-myc-tagged DN PKCδ. Cells that had been serum-starved for 1 h were stimulated with ghrelin (100 nM, 5 min). (c) CHO cells were transiently transfected with plasmid DNA encoding hGHSR-1a (10 μg) together with 20 μg of empty pEGFP-N1 vector, wild-type PKCα-GFP or c-myc-tagged DN PKCα. Cells, which had been serum-starved for 1 h, were then stimulated with ghrelin (100 nM, 5 min). Phosphorylated ERK1/2 (pERK1/2) and total ERK1/2 (ERK1/2) were detected by immunoblotting as described in Figure 2. The expression level of the different PKC isoforms was revealed by probing the western blot with anti-PKCα, δ or ɛ antibodies. All blots are representative of three experiments with similar results.