Extended Data Fig. 9: BMDM-specific SIRT2 knockout has no effect on inhibiting osteoclastogenesis and slowing down bone loss in aged and OVX mice.

a,b, Western blot analysis of SIRT2 protein expression in the primary BMDMs derived from female (a)or male (b) young (n = 3, 3 months of age) and aged (n = 3, 18 months of age) mice.c, Western blot analysis of SIRT2 protein expression in the primary BMDMs derived from sham (n = 3) and OVX (n = 3) mice (12 weeks of age). (a-c, one technical replicate of 3 biological replicates for each group). d, Western blot analysis of SIRT2 protein expression in the primary BMDMs treated with RANKL. n = 3 biologically independent experiments. (a-d, Western blot density analyzed by ImageJ and quantification analysis was shown). e-h, Represented images of 3D restoration and quantification of trabecular BV/TV, Tb.N., Tb.Sp. and Tb.Th. of distal femur of the aged female (e,f) or male (g,h) LoxP (both n = 6) and SIRT2-KO^lyz (both n = 6) mice, as measured by micro-CT. One technical replicate of 6 biological replicates for each group. i,j, Represented images of 3D restoration and quantification of trabecular BV/TV, Tb.N., Tb.Sp. and Tb.Th. of distal femur of the OVX female LoxP (n = 10) and SIRT2-KO^hep (n = 10) mice (12 weeks of age), as measured by micro-CT. One technical replicate of 10 biological replicates for each group. k,n, BMDMs were isolated from LoxP and SIRT2-KO^lyz mice (8 weeks of age) and cultured with murine M-CSF and RANKL stimulation for 7 days to generate osteoclasts. Representative TRAP staining images of osteoclasts (scale bar, 200 µm). l,m,o,p, Number and area of multi-nucleated TRAP⁺ cells of female (l-m) or male (o-p) mice. n = 3 biologically independent experiments. Data are presented as mean ± SD. with biologically individual data points shown. P values are determined by unpaired two-tailed Student’s t-test (a-d,f, h, j, l, m, o, p). n.s., not significant.

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