The type of statistical test used to analyze each outcome
| Figures | Outcome | Test | Formula |
|---|---|---|---|
| 1B | Dam mass | LMM | Mass ∼ early-life treatment × PNDa + (1|dam) |
| 1C | Dam corticosterone | t test | Corticosterone ∼ early-life treatment |
| 1D | Number of nest exits | Nonparametric longitudinal F1 LD F1 | Number of nest exits ∼ early-life treatment × PNDa |
| 1E | Percentage of time off nest | Nonparametric longitudinal F1 LD F1 | Percentage off nest ∼ early-life treatment × PNDa |
| 2A | PND4 offspring mass litter average | t test | Mass ∼ early-life treatment |
| 2B | PND11 offspring mass individual pups | LMM | Mass ∼ early-life treatment × sex + (1|dam) |
| 2C | Offspring mass males and females run separately | LMM with splines | Mass ∼ early-life treatment × PNDb + (1|dam) + (1|mouse) |
| 2D | Age at vaginal opening | LMM | Age ∼ early-life treatment + (1|dam) |
| Age at first estrus | |||
| Age at preputial separation | |||
| 2E | Mass at vaginal opening | LMM | Mass ∼ early-life treatment + (1|dam) |
| Mass at first estrus | |||
| Mass at preputial separation | |||
| Text | AGD | LMM | AGD ∼ early-life treatment × sex + (1|dam) |
| 3B | Number of estrous cycles | LMM | Number of estrous cycles ∼ early-life treatment + (1|dam) |
| 3C | Cycle length | LMM | log10(cycle length) ∼ early-life treatment + (1|dam) |
| 3D | Percentage of days in stage | χ2 test | Distribution of days spent in diestrus, proestrus, and estrus by early-life treatment |
| 4A | Male serum corticosterone | LMM | log10(cort) ∼ early-life treatment × adult treatment × time + (1|mouse) + (1|dam) |
| 4B | Female serum corticosterone | LMM | log10(cort) ∼ cycle stage × early-life treatment × adult treatment × time + (1|mouse) + (1|dam) |
| 4-1A | Body mass | LMM | Outcome ∼ early-life treatment × adult treatment + (1|dam) |
| 4-1B | Percentage of change in body mass | ||
| 4-1C | Adrenal mass | ||
| 4-1D | Adrenal mass normalized to body mass | ||
| 4-1E | Seminal vesicle mass | ||
| 4-1F | Seminal vesicle mass normalized to body mass | ||
| 4-1G | Testicular mass | ||
| 4-1H | Testicular mass normalized to body mass | ||
| 4-2A | Body mass | LMM | Outcome ∼ early-life treatment × adult treatment × cycle stage + (1|dam) |
| 4-2B | % Change body mass | ||
| 4-2C | Adrenal mass | ||
| 4-2D | Adrenal mass normalized to body mass | ||
| 4-2E | Uterine mass | ||
| 4-2F | Uterine mass normalized to body mass | ||
| 4-3B | Serum corticosterone | LMM | log10(cort) ∼ dosage × time + (1|mouse) + (1|dam) |
| 4-3A | Body mass | LMM | Outcome ∼ dosage + (1|dam) |
| 4-3C | Percentage of change in body mass | ||
| 4-3D | Adrenal mass | ||
| 4-3E | Adrenal mass normalized to body mass | ||
| 4-3F | Seminal vesicle mass | ||
| 4-3G | Seminal vesicle mass normalized to body mass | ||
| 4-3H | Testicular mass | ||
| 4-3I | Testicular mass normalized to body mass | ||
| 5A | Average LH, diestrus mice | LMM | Average LH ∼ early-life treatment × adult treatment + (1|dam) |
| 5C | Proportion with LH surge, proestrus | GLMM—logistic regression | Surged ∼ early-life treatment + adult treatment + (1|dam) |
| 6A | Capacitance | LMM | Outcome ∼ early-life treatment × adult treatment + (1|mouse) + (1|dam) |
| 6B | Input resistance | ||
| 6C | Series resistance | ||
| 6D | Holding current | ||
| 7C | PSC frequency | GLMM—NB | Number of events in 4 min ∼ early-life treatment × adult treatment + (1|mouse) (1|dam) |
| 7D | Mean interevent interval | LMM | log10(interval) ∼ early-life treatment × adult treatment + (1|mouse) + (1|dam) |
| 7E | Mean relative amplitude | LMM | Outcome ∼ early-life treatment × adult treatment + (1|mouse) + (1|dam) |
| 7F | Decay time | ||
| 7G | Distribution of interevent interval | Anderson–Darling test and bootstrapping. See Materials and Methods, Statistical analysis | |
| 7H | Distribution of amplitude | ||
LMMs were used where appropriate for the data structure and experimental design. Generalized linear mixed models (GLMM) were fit with either logistic regression or NB families.
↵a Indicates that PND was treated as a factor variable. For the analysis of dam behavior with a nonparametric longitudinal test, the nparLD package in R was used, with the F1 LD F1 Model. The subject variable was each dam, early-life treatment (STD or LBN cage) was the between-subject factor (“whole-plot” factor), and PND was the within-subject factor (“sub-plot” repeated factor).
↵b Indicates that linear splines at PND21 and 35 for the models of offspring mass allow the model to change the slope of the line for the segments between PND11 and 21, from 21 to 35, and from 35 to 72.