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. 2010 Oct 14;20(2):235–244. doi: 10.1093/hmg/ddq458

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© The Author 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Implantation of hPAE cells into the thigh muscle of BALB/c mice. (A) Human periosteal cells (2 × 10⁷ cells) were injected directly into the thigh muscles of BALB/c mice. Immunohistochemical analysis was performed on the muscle section using an antibody against vimentin. Upper panels: 2 days after injection and lower panels: 2 weeks after injection. (B) hPAE cells (2 × 10⁷ cells) were injected directly into the thigh muscles of BALB/c mice. Upper panels: immunohistochemistry against vimentin. Lower panels: immunofluorescent analysis. DAPI (blue), vimentin (green), laminin (red) and MERGE (from left to right). (C) Immunohistochemical analysis of the thigh muscle sections at 2 days or 2 weeks after injection of human periosteal cells (hPeriosteal) and at 2 weeks after injection of hPAE cells, using antibodies against vimentin (upper panels: red and lower panels: green), leukocyte marker CD45 (green) and T cell marker CD3 (red). (D) Immunofluorescent analysis using an antibody against HLA-E (red) and human laminin (green) on the thigh muscle sections at 2 weeks after injection of hPAE cells. (E) Western blot analysis of muscle lysates showing levels of HLA-E, dystrophin and laminin. BALB/c mice were implanted with PBS or hPAE cells at the indicated weeks. The level of actin protein was used as a loading control. (F) Immunofluorescent analysis using an antibody against human dystrophin (green) on thigh muscle sections 3 weeks after direct injection of hPAE cells (middle and lower panels). PBS was injected into contralateral muscles as a control (upper panels). Dystrophin is totally absent in PBS-injected muscles (upper panels), whereas clusters of muscle fibres display peripheral localization of the dystrophin protein in mice injected with hPAE cells (middle and lower panels). Dystrophin (green), DAPI (blue) and MERGE (from left to right). (G) Immunofluorescent analysis using antibodies against laminin (green), human nuclei (HuNucl, red, arrows) and DAPI staining (blue, arrowheads) on thigh muscle sections 3 weeks after injection of hPAE cells.