Extended Data Fig. 2: Liver-specific SIRT2 deficiency has no effects on osteogenic differentiation and Vitamin D synthesis.

a, Alkaline phosphatase (ALP) and alizarin red (ARS) staining after osteogenesis from bone marrow-derived mesenchymal cells (BM-MSCs) treated with LoxP-plasma and SIRT2-KO^hep-plasma for 7 days and 14 days respectively. b, Area of ALP staining and ARS staining positive osteoblasts was analyzed by ImageJ and quantification analysis is shown. c, The expression of osteogenic genes Runx2, Sp7, osteocalcin and ALP in osteoblasts as measured by real-time PCR. n = 3 biologically independent experiments. d, CYP2R1 and CYP27A1 mRNA level in livers of aged female LoxP and SIRT2-KO^hep mice (18 months of age) were analyzed by real-time PCR (aged female LoxP mice, n = 8 and aged female SIRT2-KO^hep mice, n = 9). e, Total plasma concentration of Vitamin D of aged female LoxP and SIRT2-KO^hep mice were detected by ELISA. f, CYP2R1 and CYP27A1 mRNA level in livers of aged male LoxP and SIRT2-KO^hep mice (18 months of age) were analyzed by real-time PCR (aged male LoxP mice, n = 10 and aged male SIRT2-KO^hep mice, n = 12). g, Total plasma concentration of Vitamin D of aged male LoxP and SIRT2-KO^hep mice were detected by ELISA. (d,e. one technical replicate of 8 (LoxP mice) or 9 (SIRT2-KO^hep mice) biological replicates for each group. f, g. one technical replicate of 10 (LoxP mice) or 12 (SIRT2-KO^hep mice) biological replicates for each group). Data are presented as mean ± SD. with biologically individual data points shown. P values were determined by one-way ANOVA followed by Tukey’s test (b, c). P values are determined by unpaired two-tailed Student’s t-test (d-g). n.s., not significant.

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