Figure 1: Osteoporotic phenotype of aged Nell-1-haploinsufficient mice. | Nature Communications

Figure 1: Osteoporotic phenotype of aged Nell-1-haploinsufficient mice.

From: NELL-1 in the treatment of osteoporotic bone loss

Figure 1

(a) Immunohistochemical staining for Nell-1 protein in rat spines, at 1 week, and 3 and 9 months. (b) Semiquantification of Nell-1 immunohistochemical staining, expressed as Nell-1+ bone-lining cells per B.Pm (N=20, 39 and 25 images, respectively). (c) Relative Nell-1 gene expression in mouse spines at 1 week, and 3 and 9 months, using qRT–PCR (N=3 samples per time point). (d) 3D image of liveCT and 18F radioisotope incorporation of aged (18-month old) wild-type (Nell-1+/+) and Nell-1-haploinsufficient (Nell-1+/6R) mice. (e) Quantification of BMD stratified by the lumbar vertebral level (L1–L6; N=11 and 12 mice, respectively). (f) Quantification of 18F incorporation (N=6 mice per genotype). (g) 3D coronal reconstructions of lumbar vertebrae of aged Nell-1+/+ and Nell-1+/6R mice. (h–j) Trabecular analyses of the lumbar vertebrae, including (h) trabecular bone thickness (Tb.Th), (i) number (Tb.N) and (j) spacing (Tb.Sp; N=12 and 19 vertebrae, respectively). (k–m) Serum studies among Nell-1+/+ and Nell-1+/6R aged mice including (k) serum PINP (N=19 and 17 mice), (l) serum TRAP5b (N=10 and 9 mice) and (m) serum CTX (N=12 and 22 mice). (n) Masson’s trichrome staining of lumbar vertebral bodies of Nell-1+/+ and Nell-1+/6R mice, coronal section. (o–q) Histomorphometric quantifications of lumbar vertebral bodies. Measurements include (o) B.Ar (N=29 and 26 images), (p) percentage (%) B.Ar (N=29 and 26 images) and (q) B.Pm (N=30 and 29 images). (r) TRAP staining of lumbar vertebral bodies, indicating osteoclasts. Quantification of (s) Ob.N per B.Pm (N=20 images per genotype) and (t) Oc.N per B.Pm (N=26 and 39 images). Black scale bar, 25 μm; red scale bar, 10 mm; yellow scale bar, 100 μm. HU: Hounsfield Units; pCi (pico Curie). Data points indicate the means, while error bars represent one s.e.m. Experiments were performed without replicate. Parametric data were analysed using an appropriate Student’s t-test or a one-way analysis of variance (ANOVA), followed by a post hoc Tukey’s test. Nonparametric data were analysed with a Mann–Whitney U-test or a Kruskal–Wallis one-way analysis. *P<0.05, **P<0.01.

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