Fig. 4: Satellite cell distribution in WT and MYF5/MYOD/MYF6-null porcine embryos.

Immunohistochemical staining of cross sections of hindlimbs labeled with DAPI (nuclei, blue), Laminin (basement membrane, red), and Pax7 (satellite cell, yellow). a–c Representative sections of WT hindlimbs across gestational stages E41 (a), E62 (b), and E90 (c) are displayed. d, e Representative hindlimb sections from MYF5/MYOD/MYF6-null embryos at E41 (d) and E62 (e), illustrating the lack of Pax7⁺ cells and diminished Laminin expression. Scale bar, 100 µm. f Quantification of Pax7⁺ cells per nuclei across the samples is depicted (n = 4). Bar graphs represent mean ± s.d. and statistical differences were analyzed by one-way ANOVA with Tukey’s test for multiple comparisons. (***p < 0.001). Immunohistochemical staining of cross sections of a hindlimb labeled with DAPI (nuclei, blue), Dystrophin (muscle membrane, red), and Myogenin (muscle differentiation marker, yellow). g–i Representative section of WT hindlimb skeletal muscle across developmental stages E41 (g), E62 (h), and E90 (i) are displayed. j, k Representative sections from MYF5/MYOD/MYF6-null hindlimbs at E41 (j) and E62 (k), showing the absence of Myogenin⁺ cells and Dystrophin. Scale bar, 100 µm. l Myogenin⁺ cells quantification across the samples is presented (n = 4). Bar graphs represent mean ± s.d. with statistical validation using one-way ANOVA and Tukey’s test (**p < 0.01, ***p < 0.001).