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. 2023 Nov 1;14:6993. doi: 10.1038/s41467-023-42837-8

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© The Author(s) 2023

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. 2 — a Scheme of the experimental strategy. b Representative profiles of fluorescence-activated cell sorter analysis of the percentage of Gli1⁺ MuSCs from Gli1-CreERT2; R26-tdT mice at different time points with oil or TAM treatment (n = 5). tdT-marked Gli1⁺ MuSCs were detected within 3 days and plateaued at day 14, showing no further increase at day 30. c Quantification analysis of the percentage of MuSCs expressing tdT at different time points with oil or TAM treatment (n = 5). d Scheme of the experimental strategy for analyzing TAM-induced Gli1⁺ MuSCs. e Immunofluorescence staining for tdT (red), Pax7 (green), Laminin (gray) and DAPI (blue) in TA muscles (n = 6). Scale bars, 50 µm. Boxed regions are magnified on the low. f Quantification of the percentage of Pax7⁺ cells expressing tdT (Gli1) (n = 6). g Immunofluorescence staining for tdT (red), Pax7 (green) and DAPI (blue) in isolated single EDL myofibers from hindlimb skeletal muscles (n = 5). Scale bars, 10 µm. h Scheme of the dual-recombinase genetic lineage-tracing strategy. i Schematic of the experimental strategy. j Immunofluorescence staining for tdT (red), Pax7 (green) and DAPI (blue) in TA muscle sections. White arrowhead indicated the tdT⁺ (Gli1⁺) Pax7⁺ cells from different mouse lines (n = 5). White arrows indicated the tdT⁻ (Gli1⁻) Pax7⁺ cells. Scale bars, 50 µm. k Quantification of the percentage of Pax7⁺ cells expressing tdT (Gli1) in different mouse lines (n = 5). l The model specific of MuSC populations. Red indicated the Gli1⁺ MuSCs. Green indicated Gli1− MuSCs. Light red indicated myofibers. Data are presented as mean ± SEM; Statistical significance was determined by one-way ANOVA (c, k). All numbers (n) are biologically independent experiments. Source data are provided in the Source Data File.