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Ligand-dependent Ret activation limits the trophic activity of GDNF on sympathetic neurons. The metabolic activity of neurons was determined using an MTT reduction assay as described in Materials and Methods. Neurons were treated with NGF (50 ng/ml), GDNF (50 ng/ml), epoxomicin (Epox; 5 μm), or GDNF in the presence of epoxomicin for 4 h. MTT reduction was then measured and graphed as a percentage of neurons treated with medium alone (No Treat). Error bars represent SEM.
