Supplementary Figure 6: E2 normally induces Mib1 in MFs lacking Ar.

(a,b) The Cre-mediated recombination was confirmed by PCR in Ar^ΔMF mice at the indicated ages in the shown organs (a) and in myofiber but not in satellite cells (b). Unprocessed original scans of blots are shown in Supplementary Fig. 10. (c) The gross morphology in levator ani (LA) muscles from 4-week-old Ar^ΔMF mice. Scale bars, 0.5 cm. (d,e) IHC staining (d; scale bars, 20 μm), and quantification (e; n = 3 animals for each group; data are mean ± s.d.; Bootstrap t-test; ^∗P < 0.05) for Pax7, Ki67, and Laminin in TA muscles from 10-day-old Ar^WT (upper) and Ar^ΔMF (bottom) mice injected with Veh or E2. TA muscles were isolated from hindlimb muscles 3 days after treatment. Arrows and arrowheads indicate Ki67⁺Pax7⁺ and Ki67⁻Pax7⁺ cells, respectively. Statistical source data for e is provided in Supplementary Table 2. (f,g) Muscle mass (f; n = 3 animals for each group; data are mean ± s.d.; two-sample t-test; ^∗P < 0.05), myonuclei numbers in MFs (g; n = 3 animals for each group; data are mean ± s.d.; Poisson’s general linear model regression; N.S. not significant) of 12-week-old Cont^WT and Ar^ΔMFEsr2^−/− mice. Muscle mass, myonuclei numbers were comparable. Statistical source data for f and g is provided in Supplementary Table 2. (h) Mib1 levels in 12-week-old EDL muscles from indicated genotypes. n = 3 animals for each group; data are mean ± s.d.; Tukey’s pairwise comparison test; ^∗P < 0.05. (i–k) IHC staining for Mib1 (i; scale bar, 100 μm), the expression of Ar and Esr2 (j) in MFs, and the mRNA expression of Notch target genes in MCs (k). n = 3 animals for each group; data are mean ± s.d.; two-sample t-test; ^∗P < 0.05. Statistical source data for j and k is provided in Supplementary Table 2. The MFs were isolated from hindlimb muscles of 12-week-old Cont^WT and Ar^ΔMFEsr2^−/− mice.