Statistical analysis
Survival curves were generated using the Kaplan–Meier method, and survival was compared using the log-rank test. Statistical comparisons between two groups and among three groups were conducted using Student’s t test and one-way analysis of variance, followed by a Bonferroni post hoc test, respectively. Data are presented as mean ± standard deviation (SD). All statistical analyses were performed using JMP 14. software (SAS Institute Inc., Tokyo, Japan). p values <0.05 were considered statistically significant. Normality testing was performed using the above statistical analysis.
Based on the hypothesis for the primary study endpoint of survival rates, a sample size calculation was performed using G*Power, version 328. The effect size decided to be clinically significant was 0.6. We permitted a type I error of α=0.05, and with the alternate hypothesis, the null hypothesis would be retained with a type II error of β=0.2. Sample size analysis resulted in the need for 30 animals. The final sample size for each group complies with the ARRIVE guidelines22.