Fig. 4

SIRT3 is reduced in HFD&STZ mice and HGPA-treated osteoblasts. a Heatmap demonstration of Sirtuin family (Sirt1-7) and expression of Sirt3 in bone from CTRL and HFD&STZ mice. b IHC staining of SIRT3 in bone sections. c, d IF analysis of bone sections showing co-localization of SIRT3 with osteoblast marker (OCN) or osteoclast marker (CTSK). Scale bar, 100 μm. e, f Western blot analysis of SIRT3 in primary osteoblasts and osteoclasts from CTRL and HFD&STZ mice. g, h Quantification of western blot and qPCR analysis of Sirt3 gene expression in osteoblasts and osteoclasts from CTRL and HFD&STZ mice. i Western blot analysis and quantification of SIRT3 expression in bone tissue from CTRL and HFD&STZ mice. j–m Western blot analysis and quantification of SIRT3 protein in MC3T3-E1 cells cultured for different time and concentration of HGPA. Data presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001