The data were statistically analyzed with Prism 8 (GraphPad Software, La Jolla CA, USA). All data were expressed as mean ± standard deviation (SD). The sample sizes used in each experiment were based on our previous studies16 (link),17 (link) and recent literature.24
A power analysis was performed to detect significant differences with minimal animal numbers based on our experimental experience with 80% power. Therefore, n = 5–10 was chosen. All data were included in the analyses. The data from the behavioral test were analyzed with two-way repeated-measures analysis of variance (RM ANOVA) followed by Tukey’s or Bonferroni’s post hoc test. Western blot data was compared using two-tailed t tests. A value P < 0.05 was considered significant. Image-Pro Plus analysis software (Media Cybernetics, Silver Spring, MD, USA) was used to measure the density of specific bands from western blotting. The results were shown by the ratio of phospho-µ-opioid receptor to β-actin immunoreactivity.
A power analysis was performed to detect significant differences with minimal animal numbers based on our experimental experience with 80% power. Therefore, n = 5–10 was chosen. All data were included in the analyses. The data from the behavioral test were analyzed with two-way repeated-measures analysis of variance (RM ANOVA) followed by Tukey’s or Bonferroni’s post hoc test. Western blot data was compared using two-tailed t tests. A value P < 0.05 was considered significant. Image-Pro Plus analysis software (Media Cybernetics, Silver Spring, MD, USA) was used to measure the density of specific bands from western blotting. The results were shown by the ratio of phospho-µ-opioid receptor to β-actin immunoreactivity.
