Fig. 7.

Administration of KDM4B inhibitors ameliorates bone loss in the ovariectomized (OVX) mouse model. a TRAP staining of BMMs treated with ML324. Scale bar, 75 μm. b mRNA expression levels of Fosl2 and Tpm1 in the cells evaluated in (a). c BMMs were treated with vehicle or 10 μmol·L⁻¹ ML324 in the presence or absence of RANKL (100 ng·mL⁻¹) for 10 days. The dentin slices were stained with Mayer’s hematoxylin, the resorption pit area was analyzed. Scale bar, 200 μm. d, e ChIP assay of H3K9me3, KDM4B, CCAR1, and p65 occupancy at the Fosl2 promoter or Tpm1 promoter in BMMs treated with ML324 (10 μmol·L⁻¹) and/or RANKL for 30 min. f Micro-CT analysis of the femurs of 4-month-old sham-operated, saline-treated (Sham), saline-treated OVX (OVX), and ML324-treated OVX (OVX + M 1.74) mice (n = 5). Left panel, representative micro-CT image of a proximal femur (top, axial view; bottom, longitudinal view). Right panel, bone parameters. M 0.35, 0.35 mg·kg⁻¹ body weight; M 1.74, 1.74 mg·kg⁻¹ body weight. Scale bars, 1 mm. g Histological analysis of femur sections from the mice evaluated in (f) (n = 3). ML324, 1.74 mg·kg⁻¹ body weight. The sections were stained for H&E and TRAP. The arrows indicate osteoclasts. Scale bars, 100 μm. The data are presented as the mean ± SD values of three independent experiments [one-way ANOVA in (a–c) and two-way ANOVA in (d, e)]. The data in (f, g) are presented as the mean ± SEM values (one-way ANOVA). *P < 0.05; **P < 0.01; ***P < 0.001. See also Supplementary Figs. 7 and 8