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. 2021 Feb 26;12:1318. doi: 10.1038/s41467-021-21631-4

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© The Author(s) 2021, corrected publication 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Upper panels: Immunofluorescence analysis of muscle stem cells associated with fibers from mice carrying the Dll1^luc allele; myofibers were freshly isolated (0 h), or cultured for 24 and 72 h as indicated. Lower Panels: corresponding differential interference contrast (DIC) images. Dll1-Luc (anti-luciferase; red), Pax7 or MyoG (blue), MyoD (green); n = 4 animals. b RNAscope analysis of a regenerating tibialis anterior (TA) muscle at 4 days post injury (dpi); MyoD or MyoG (red), Dll1 (blue), Pax7 (green); DAPI was used as counter stain (white); n = 5 animals. c Quantification of colony size of muscle stem cells on fibers from control (TxCon; blue bars) and Dll1 mutant (TxDll1^f/f; red bars) animals; fibers were cultured for 60 and 72 h. n = 3 animals. d Quantification of cells that express Pax7 (red bars), Pax7 and MyoG (yellow bars), MyoG (green bars) in colonies associated with myofibers from control (TxCon) and Dll1 null mutant (TxDll1^f/f) animals cultured for 72 h; n = 3 animals. e Schematic outline of the regeneration experiment shown in f,g. The Dll1 null mutation was induced by tamoxifen (TxDll1^f/f mutant mice); black arrows: tamoxifen injections; green arrow: cardiotoxin (CTX) treatment; red arrows: analysis of muscle regeneration. f Immunohistological analysis of cells expressing Pax7 (red) and MyoG (green) in the injured muscle of control (TxCon) and Dll1 null mutant (TxDll1^f/f) animals at 4dpi (left panels). Quantifications of Pax7+ and MyoG+ cells (right panels). Quantified were the numbers of Pax7+ or MyoG+ cells/mm² in TxCon (blue bars) and TxDll1^f/f (red bars) animals, which is also displayed as the proportion of Pax7 + (red) and MyoG + (green) cells (n = 3 animals). g Immunohistological analysis of cells expressing Pax7 (green), Collagen IV (ColIV, blue) in the injured muscle of control (TxCon) and Dll1 null mutant (TxDll1^f/f) animals at 7dpi (left) and 21 dpi (right); DAPI (red) is used as counterstain. Quantifications of Pax7+ cells/mm², nuclei/fiber and fiber diameter are shown below (TxCon, blue bars and TxDll1^f/f, red bars); n = 3 animals. Scale bars, 10 μm (a, b) and 50 μm (f,g). Data are presented as mean values ± SEM. Exact p values are indicated, unpaired two-sided t-test.