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Figure 2. Effect of pharmacological inhibitors on leptin-mediated phosphorylation of ATF-1 and expression of neurotensin gene expression. A) Cells were plated in 60 mm plates to 
80% confluence, then serum-starved overnight. After pretreatment with or without 10 µM SB 203580 for 1 h, N-39 neurons were stimulated with leptin (10–11 or 10–7 M, respectively) for 15 min. Cell lysates were examined by Western blot with phosphospecific and total CREB antibodies. A summary of all experiments performed (n
3) is presented in the graph as mean ± SEM; *significantly different (P<0.05) from untreated control. Immunoblot is representative of an experiment performed at least 3 times. B) Cells were plated in 60 mm plates to 80% confluence, then serum-starved 2 h. After pretreatment with or without the specified pharmacological inhibitors for 1 h, N-39 neurons were stimulated with leptin (10–11 or 10–7 M) for 4 h. Neurotensin mRNA expression was determined by real-time PCR. Values for NT are expressed relative to 
-actin mRNA levels (mean±SE, n=3). *Significantly different (P<0.05) compared with untreated control.
