Bone Resorption Assay of BMMs

Bone resorption was assessed by the bone resorption assay kit (Cosmo Bio Co., Ltd., Tokyo, Japan), as previously described [45 (link)]. The BMMs were seeded into a 48-well bone resorption assay plate at a density of 2.5 × 10⁴ cells/well. The cells were differentiated with M-CSF and RANKL in the absence or presence of indicated concentrations of LSD1 inhibitors. After cell differentiation, the attached cells were removed using 5% sodium hypochlorite. The plates were air-dry at RT and resorption pits were captured by a microscope. In addition, the resorption areas were quantified by analyzing three randomly selected pictures per well by Image J software.