KDM6B orchestrated the polarization of BMDMs
and the osteogenic
differentiation of BMSCs by regulating BMAL1 expression. (a, b) BMDMs
were isolated from the sham surgery and OVX cohorts, and RT-qPCR and
WB assay were performed to detect the effect of KDM6B and GSK-J4.
(c, d) BMAL1-KO BMDMs and BMAL1-OE BMDMs were isolated from BMAL1-KO
mice and lentivirus-infected BMDMs, respectively. RT-qPCR and the
WB assay were performed for subsequent analyses. (e–g) Flow
cytometry and immunofluorescence were used to evaluate the results
of macrophage polarization in BMAL1 knockout and overexpression systems
(scale bar: 100 μm). (h) BMSCs were seeded in the lower chamber
of the Transwell plate, while BMDMs with different treatments were
added to the upper chamber. Analysis of osteogenic differentiation
of BMSCs in the lower chamber was completed by ALP staining or Alizarin
Red S staining (scale bar: 100 μm). (i) The WB results show
the expression of osteogenesis-related genes.