Fig 1: Decreased MEN1 expression in fibrotic kidney disease samples. (A) Quantitative real-time PCR (qPCR) was used to detect the mRNA expression of Men1, Acta2, Fibronectin1, Col1a1, Col3a1 and ECM2 in the kidney tissues of the sham and unilateral ureteral obstruction (UUO) mice (n = 8 mice per group). (B) Representative images of haematoxylin–eosin (H&E), Masson's trichrome and Sirius red staining of kidney sections from the sham and UUO mice; scale bars 50 µm. (C) Quantification of the interstitial fibrosis score, the area of Masson's trichrome and Sirius red staining in the kidney tissues of the sham and UUO mice (n = 8 mice per group). (D) Immunohistochemistry (IHC) staining for menin, a-SMA and collagen 1 in the kidney tissues of the in the kidney tissues of the sham and UUO mice; scale bars 50 µm. (E) Quantification of menin, a-SMA and collagen 1 IHC staining in (D) (n = 8 mice per group). (F) Western blotting was used to detect the expression of the indicated proteins in the kidney tissues of the sham and UUO mice (n = 3 mice per group). (G) Western blotting was used to detect the expression of the indicated proteins in mouse renal tubular epithelial cells (mRTECs) at the indicated time points after exposure to 10-ng/ml TGF-ß. (H) Representative images of menin and H3K4me3 IHC staining in kidney tissues of the minimal change diseases (MCD) and diabetic nephropathy (DN) patients; scale bars 50 µm. The data are represented as the mean ± standard deviation (SD); *p < .05, **p < .01, ***p < .001, ****p < .0001.
Fig 2: Effects of D2 on bone volume and its mechanical properties in a mouse model of osteogenesis imperfecta (OI). (A) Comparison of postnatal growth (left) and gross appearance (right) between wild-type (WT) and Mov13 male mice. *p < 0.05 (Student's t-test). (B) Bone volume of L3 vertebral bodies of WT and Mov13 female mice orally administered D2 (10 mg/kg/day) or vehicle. 3D micro-CT images (left) and BV/TV (right) of the L3 vertebrae are shown for WT and Mov13 female mice at the age of 8 weeks (after 4-week D2 administration to Mov13 mice). Data are means ± SD (n = 4 to 5). *p < 0.05 (Steel–Dwass test). (C) Mechanical properties of L3 vertebral bodies of WT and Mov13 female mice orally administered D2 or vehicle. Load and deformation were measured by compression tests (left). The maximum load is shown in the right panel. Data are means ± SDs (n = 11). *p < 0.05 (Steel–Dwass test). (D) Production of type I collagen in primary osteoblasts isolated from either WT or Mov13 mouse calvaria at P1. Cells were treated with or without 10 nM D2 for 2 days. The cellular production of type I collagen was measured by ELISA. Data are means ± SDs (n = 6). *p < 0.05, **p < 0.01 (Steel–Dwass test).
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