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Fig. 6 Musk ambrette is a candidate kisspeptin receptor (KISS1R) agonist. A, Gonadotropin-releasing hormone receptor (Gnrh1) expression levels (determined with quantitative reverse-transcription polymerase chain reaction [qRT-PCR]) in murine hypothalamic cells after various concentrations of musk ambrette treatment at 1, 2, 4, or 6 hours. B, Gnrh1 expression levels (determined with qRT-PCR) in murine hypothalamic mHypo-GnRH/GFP spheroids after various concentrations of musk ambrette at 1 hour. C, mHypo-GnRH/GFP spheroid morphology at 6 days pre treatment. There was no change in morphology after treatment (data not shown). Spheroids continued to appear as uniform, solid cell aggregates with a spherical shape and a diameter of approximately 200 to 250 μm. D, GNRH1 expression levels (determined with qRT-PCR) in induced pluripotent stem cells–derived human hypothalamic neurons after 25 μM musk ambrette treatment at 1, 2, 4, or 6 hours. E, Molecular docking predicts the potential binding sites between musk ambrette and KISS1R. All values represent the mean ± SD (n = 3 replicates). Statistical analysis was performed using a 2-way analysis of variance with Tukey multiple-comparison test, and the P value is shown in each graph.