Data and statistical analysis
Data collection and statistical analysis comply with the recommendations
of the British Journal of Pharmacology on experimental design and
analysis in pharmacology (Curtis et al., 2018). The number of
experiments in each experimental group was selected on the basis of
observing a statistically significant effect while using the minimum
number of animals (3R principles) and on the experience from the
previous studies. Animals were randomly assigned in the different
experimental groups with each group having the same number of animals by
design. Blinding of the operator was not feasible, but data analysis was
performed semi-blinded by an independent analyst.
Statistical analysis was performed using Statistica 6.1 Base (Tulsa, OK,
USA). Shapiro–Wilk normality test was used to analyze the data
distribution. Null-hypothesis testing was performed by ANOVA. One-way
ANOVA followed by Dunnett’s or Tukey’s test for multiple comparison post
hoc was used. For related groups one-way repeated ANOVA test was
performed. Data are presented as mean ± SEM. Values of P <
0.05 were considered significant.