Figure 2. Bethanechol and E2 induce ERK phosphorylation and NO production in TM-BBB cells in a time-dependent manner.

(A) TM-BBB cells were stimulated with 100 µM of the muscarinic agonist, bethanechol (Bch), or 10 nM E2 for different periods of time (1–60 min), and cell lysates were analyzed by western blotting with anti-pERK or anti-total ERK antibodies. The cumulative results of Western blot analysis are reported from three independent experiments (10 min after treatment with Bch or E2). Values are means±SEM. *p<0.05 (vs vehicle control). (B) TM-BBB cells were stimulated with 100 µM Bch or 10 nM E2 for different time periods. Subsequently, nitrite levels were measured in the culture medium. Values are means±SEM. **p<0.001 (vs vehicle control).