Endothelin-1 regulates cardiac sympathetic innervation in the rodent heart by controlling nerve growth factor expression
J. Clin. Invest. Masaki Ieda, et al. 113:876 doi:10.1172/JCI19480 [Go to this article.]

Figure 3
ET-1 causes NGF-mediated differentiation of PC12 cells. (A) PC12 cell morphology was observed after incubation for 3 days in mock medium or medium conditioned with unstimulated cardiomyocytes (U-CM) or ET-1–stimulated cardiomyocytes (ET-CM). Medium conditioned with ET-1–stimulated cardiomyocytes strongly induced neurite extension in PC12 cells compared with medium conditioned with unstimulated cardiomyocytes. (B) Percentage of differentiated cells in A (n = 4). (C) PC12 cells were pretreated with anti-NGF blocking antibody for 30 minutes, then incubated with the conditioned medium (n = 4). (D) PC12 cells transfected with LacZ were cocultured with cardiomyocytes, then stimulated with ET-1 or ET-1 plus anti-NGF blocking antibody for 3 days. PC12 cells were identified using X-gal staining. (E) Percentage of differentiated cells in D (n = 4). (F) NGF protein levels in medium conditioned with Edn1^+/+ or Edn1^–/– cardiomyocytes were measured by ELISA (n = 4). *P < 0.0001; **P < 0.001; ^#P < 0.01. Scale bar: 100 μm.