The results obtained were expressed as the mean and standard error of the mean (SEM). These results were analyzed statistically using Student's t-test or two-way analysis of variance (ANOVA) followed by Bonferroni's post-test, and the differences between means were considered significant when p < 0.05. The pCE50 values were calculated by non-linear regression (Neubig et al., 2003). The maximum effect (Emax) values were obtained by averaging maximum percentages of contraction or relaxation. All results were analyzed by GraphPad Prism® version 5.01 (GraphPad Software Inc., San Diego, CA, USA).
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