Fig. 1

ZEB1 is upregulated in dystrophic muscles and is expressed by undamaged myofibers. a Zeb1 mRNA levels in the gastrocnemius muscles of 2-month-old wild-type and mdx mice were assessed by qRT-PCR. Data are the average of six mice for each genotype. Throughout the Figures, relative data in percentage are shown with the value of the wild-type or control condition arbitrarily set to 100. b The gastrocnemius muscles of wild-type and mdx mice were assessed for ZEB1 (clone H102) and laminin (4H8-2) along with DAPI for nuclear staining. Representative peripheral and centralized nuclei were labeled with arrows and arrowheads, respectively. For mdx muscles, two different areas are shown: one that predominantly exhibits damaged fibers (upper panel), and another with signs of regeneration (lower panel). See Supplementary Fig. 1A for individual staining captures. Scale bars: 25 μm (wild-type mice) 50 μm (mdx mice). c The percentage of ZEB1⁺ peripheral nuclei in b was calculated either out of the total number of nuclei (peripheral plus centralized) or only with respect to peripheral nuclei. Data are the mean of at least five fields from three mice for each genotype. d As in b, but 9–12 h before euthanasia mice were injected with EBD. See Supplementary Fig. 1C for individual staining. Scale bar: 50 μm. e Human healthy and dystrophic muscles were stained for ZEB1 (HPA027524) and laminin (4H8-2) along with DAPI. Representative peripheral and centralized nuclei were labeled with arrows and arrowheads, respectively. A representative area with immune cell infiltration is labeled with an asterisk (“*”). See Supplementary Fig. 1D for single staining captures. Scale bar: 50 μm. f Correlation between ZEB1 expression and CK levels in dystrophic human muscles. g Relative number of fibers expressing ZEB1 in human dystrophic muscles with respect to their CK levels below or above the median. See Supplementary Fig. 1E for representative scores of ZEB1 staining. h Left panel: Gastrocnemius muscle lysates from 2-month-old wild-type and Zeb1 (+/−) mice (two per genotype, labeled as 1 and 2) were blotted for ZEB1 (HPA027524) and GAPDH (14C10) as loading control. Right panel: As in the left panel but from 2-month-old mdx;Zeb1 (+/+) and mdx;Zeb1 (+/−) mice, three for each genotype. See Supplementary Fig. 1F for full unedited blots. i Zeb1 mRNA levels in the gastrocnemius of the four genotypes were determined by qRT-PCR. Data are the average of six mice per genotype. j Wild-type and Zeb1 (+/−) gastrocnemius muscles were either counterstained with hematoxylin/eosin (H&E) (upper panel) or immunostained for laminin (4H8-2) and DAPI (lower panel). Scale bars: 50 and 100 μm, respectively