"On PND 56, 15 animals from 5 litters were killed for blood collection and anogenital distance (AGD) measurement, and then testes and cauda epididymis were collected for further analysis."

"Histological examination and IF staining are described in Supporting Information. Based on IF images, the germ cell number per seminiferous tubule as well as the numbers of spermatocytes and Leydig cells per 200 × field were counted using ImageJ software (National Institutes of Health, USA; version 1.53c). Based on the images of LAMININ1 IF for the lamina of the seminiferous tubule, the tubule area was measured. Staging of seminiferous epithelium cycle was performed based on PAS-stained sections."

"For seminiferous tubule area and germ cell number per tubule, at least 100 seminiferous tubule cross sections per testis and 5 litters per group were used for statistical analysis. For spermatocyte number and Leydig cell number, at least three 200×feld per testis and five litters per group were used for statistical analysis."

"Statistical analysis

Data are shown as mean ± standard deviation (SD). Data were analyzed by SPSS (version 16.0) or GraphPad Prism 8 software. If homogeneity, data were analyzed by independent t test (between two groups) or one-way ANOVA (treatment groups relative to the control group) followed by Duncan (equal variances assumed) and Dennett’s T3 (equal variances not assumed). When samples were heterogeneous, nonparametric analysis of variance was applied. Testis weight was analyzed using analysis of covariance with the body weight as a covariate. A p < 0.05 was considered statistically significant."

F1 Male CD-1 Mice (pre- and postnatal exposure, evaluated PND56)