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. 2021 Dec 6;15:780698. doi: 10.3389/fnins.2021.780698

FIGURE 2.

FIGURE 2

Pituitary gonadotropin serum levels and subunit gene expression in female mice exposed to CB NPs. CB NPs (100 μg) or control saline solution (NaCl, 0.9%) were intratracheally administered to C57BL/6 mice once a week during four weeks as described in the Materials and Methods. Mice were sacrificed and anterior pituitaries and blood were collected. Concentrations of FSH and LH were determined in serum by Luminex technology in 24 NaCl- and 33 CB NP-treated mice (A). Fshb, Lhb and Cga mRNA levels were determined by real-time quantitative PCR in 24 NaCl- and 33 CB NP-treated mice (B). Serum levels of inhibin B and sex steroids (estradiol and progesterone) were determined by ELISA and GC-MS, respectively, as described in the materials and methods (C). Assays were performed on 23 NaCl- and 30 CB NP-treated mice for inhibin-B, 11 NaCl- and 18 CB NP-treated mice for estradiol and 16 NaCl- and 24 CB NP-treated mice for progesterone. Data are represented as mean ± SEM. Statistical differences were determined by the Unpaired t-test (for B, Fshb mRNA) or Mann-Whitney test. *p < 0.05 and **p < 0.01 compared to NaCl.