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. 2023 Jan 30;17(3):3153–3167. doi: 10.1021/acsnano.2c12440

Figure 6.

Figure 6

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.