Figure 6

Effect of L-NAME (500 μM) on UTP (100 μM), norepinephrine (NE, 100 μM), NGF (100 ng/ml) and EGF(100 ng/ml) induced ERK phosphorylation in PC12 cells. HFα_1A-transfected PC12 cells were serum-starved for 2 hr, pretreated with L-NAME for 30 min, and exposed to agonist for 15 min before harvesting. ERK activation was determined by Western blotting with dual phospho-specific ERK antibodies as described in Methods. Although the blot shown is overexposed, similar strength of signals were seen with NE, NGF and EGF after shorter exposures.