"On PND 9, testes of male pups were collected (n=3 litters including 9 male pups) for further analysis. In addition, hypothalamus tissue was also collected for analysis for GnRH content and Gnrh expression, given crucial roles of GnRH in the production of luteinizing hormone (LH) and testosterone and subsequent testicular development and spermatogenesis."

"Measurement of the testosterone, LH, FSH, and GnRH

The content of GnRH (Qisong Bio, QS43291) in the tissue homogenate of the hypothalamus on PND 9 and levels of serum testosterone (Cayman Chemical, Cat NO. 582701), LH (Qisong Bio, QS43318), follicle-stimulating hormone, follicle-stimulating hormone (FSH) (Qisong Bio, QS43242) on PND 56 were determined by the enzymelinked immunosorbent assay (ELISA) according to the manufacture’s protocol. All the samples were analyzed in the same assay in duplicates."

"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 (postnatal exposure, evaluated PND9)