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Fig. 3 Impaired muscle regeneration and the potential aging mechanisms in the skeletal muscle of slc39a11 Mut zebrafish. (A) Schematic diagram depicting the protocol for inducing unilateral cryoinjury. The cryoprobe was precooled in liquid nitrogen and immediately placed on the right side of the anesthetized zebrafish for 6 s; 14 days after cryoinjury (14 dpci), samples were collected and analyzed. (B) Schematic diagram depicting a coronal section through the caudal peduncle of a cryoinjured sample. (C and D) Summary of myod1, myog, and p62 mRNA measured in the cryoinjured muscle tissue of male and female WT and Mut zebrafish. (E) Representative images showing the pattern of regenerated myomeres in male and female WT and Mut zebrafish. (F and G) Quantification of the gap (interspace) percentage (left panels) and relative length (right panels) of muscle fibers in the uninjured and cryoinjured sides in myomere samples obtained from male (F) and female (G) WT and Mut zebrafish. (H and I) Significant KEGG pathways related to the aging process in male (H) and female (I) groups that were enriched in Mut samples compared to WT samples, based on GSEA enrichment analysis. (J) Heatmap of the relative change in the expression of the indicated muscle regeneration-related marker genes in male and female WT and Mut samples. (K) Bubble chart analysis of the expression of the indicated muscle regeneration-related marker genes in male and female WT and Mut samples. (L) Volcano map of the expression of aging-related marker genes. (M) Fluorescence images of γ-H2AX immunostaining (green) in muscle fibers in male and female WT and Mut zebrafish; the nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.