Supplementary Figure 8: Genetically disturbed hypothalamic and gonadal axis impairs the establishment of SC population.

(a,c) Gross morphology of seminal vesicles, testes and LA muscles (a; scale bar, 0.2 cm), muscle mass (b; n = 3 animals for each group; data are mean ± s.d.; Two-sample t-test; *P < 0.05) and myonuclei number of EDL muscles (c; n = 3 animals for each group; data are mean ± s.d.; Poisson’s general linear model regression; N.S. not significant) in 8-week-old Gnrh1^+/+ and Gnrh1^hpg/hpg mice. Muscle mass and myonuclei numbers were comparable in Gnrh1^+/+ and Gnrh1^hpg/hpg mice. (d,e) The expression of Ar and Esr2 in MFs (d) and Notch target genes in SCs (e). n = 3 animals for each group; data are mean ± s.d.; two-sample t-test; ^∗P < 0.05; ^∗∗P < 0.01. (f) IHC staining (f) for Pax7 and Laminin in TA muscles. MFs, SCs, and TA muscles were isolated from 8-week-old Gnrh1^+/+ and Gnrh1^hpg/hpg mice. Scale bar, 50 μm. (g) Flow cytometric analysis of mononuclear cells from 8-week-old control (sum data of Sham, Veh, and Gnrh1^+/+), Orx, Antide-treated, and Gnrh1^hpg/hpg hindlimb muscles. n = 3 animals for each group; data are mean ± s.d.; Bootstrap t-test; ^∗P < 0.05. (h) IHC staining of Pax7 in Vcam-1⁺Sca-1⁻ cells sorted from mononuclear cells of Gnrh1^+/+and Gnrh1^hpg/hpg mice. Scale bar, 50 μm. (i) The representative chromatograms of E2 measurements by LC-MS/MS. n = 3 animals for each group; Data are mean ± s.d.