Volume 78, Issue 6, Pages (September 2010)

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Volume 78, Issue 6, Pages 561-568 (September 2010) Rapamycin induces growth retardation by disrupting angiogenesis in the growth plate Óscar Álvarez-García, Enrique García-López, Vanessa Loredo, Helena Gil-Peña, Julián Rodríguez-Suárez, Flor Á. Ordóñez, Eduardo Carbajo-Pérez, Fernando Santos Kidney International Volume 78, Issue 6, Pages 561-568 (September 2010) DOI: 10.1038/ki.2010.173 Copyright © 2010 International Society of Nephrology Terms and Conditions

Figure 1 Collagen X (Col X) expression and alkaline phosphatase in the growth cartilage of control and rapamycin-treated rats (RAPA). Representative images of in situ hybridization (a) and immunohistochemistry (b) of collagen X and alkaline phosphatase stain (c) are shown. Bar=100μm. Kidney International 2010 78, 561-568DOI: (10.1038/ki.2010.173) Copyright © 2010 International Society of Nephrology Terms and Conditions

Figure 2 In situ hybridization and immunohistochemistry analysis in the growth cartilage of proximal tibiae of control and rapamycin-treated rats (RAPA). Representative images of in situ hybridization experiments for vascular endothelial growth factor (VEGF) (a), insulin-like growth factor I (IGF-I) (c), IGF-I receptor (IGF-IR) (e), and growth hormone receptor (GHR) (g) mRNAs, and of immunohistochemistry experiments for VEGF (b), IGF-I (d), and IGF-IR (f) are shown. Bar=100μm. Kidney International 2010 78, 561-568DOI: (10.1038/ki.2010.173) Copyright © 2010 International Society of Nephrology Terms and Conditions

Figure 3 Quantitative PCR experiments. (a) mRNA expression of vascular endothelial growth factor (VEGF) and HIF1α, in the whole kidney and the growth plate of control rats (white columns) and rats treated with rapamycin (black columns). (b) mRNA expression of insulin-like growth factor I (IGF-I), IGF-I receptor (IGF-IR), and growth hormone receptor (GHR) in the liver and the growth plate of control rats (white columns) and rats treated with rapamycin (black columns). Values are expressed as mean±s.e.m. of gene fold-change relative to control group and normalized by glyceraldehyde 3-phosphate expression levels. aMeans statistically different from C group (P≤0.05). Kidney International 2010 78, 561-568DOI: (10.1038/ki.2010.173) Copyright © 2010 International Society of Nephrology Terms and Conditions

Figure 4 Western blot experiments. (a) Analysis of vascular endothelial growth factor (VEGF) in the growth cartilage and kidney of control and rapamycin-treated rats (RAPA). (b) Analysis of LC3-I (18kDa) and LC3-II (16kDa) in the growth cartilage and liver of control (white columns) and rapamycin-treated rats (RAPA, black columns). Data are representative of three blots giving similar results. aMeans statistically different from control group (P≤0.05). GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Kidney International 2010 78, 561-568DOI: (10.1038/ki.2010.173) Copyright © 2010 International Society of Nephrology Terms and Conditions

Figure 5 Immunohistochemistry analysis of insulin-like growth factor binding proteins (IGFBPs) in the growth cartilage of proximal tibiae of control and rapamycin-treated rats (RAPA). Representative images of immunohistochemistry experiments for IGFBP-3 (a) and IGFBP-5 (b) are shown. Bar=100μm. Kidney International 2010 78, 561-568DOI: (10.1038/ki.2010.173) Copyright © 2010 International Society of Nephrology Terms and Conditions