Ccn6 depletion causes loss of muscle mitochondrial abundance and alterations in muscle organization. (A) Immunofluorescence of the muscle sections of ccn6 morpholino-injected and uninjected zebrafish using anti-Vdac1 antibody with Alexa Fluor 546 rabbit secondary antibody, demonstrating loss of Vdac1 (mitochondrial structural protein) expression upon Ccn6 depletion: representation of three different experimental sets. Arrow marks denote the presence of mitochondria along the muscle fiber edges near the sarcolemma and in patches in the inter-myofibril spaces. (B) Immunoblot of the muscle lysate and corresponding densitometry showing a reduction in Vdac1 levels in ccn6 morpholino, but not control morpholino-injected fish, as compared to the uninjected control. β-actin is used as a reference protein for this estimation. Data are presented from two independent experiments; tissue was pooled from 10 fish in each experiment. (C) Histology (hematoxylin and eosin staining) of the muscle sections from ccn6 morpholino/control morpholino-injected and uninjected zebrafish demonstrating an increase in the interstitial gaps and presence of inflammatory infiltrates (arrow marks) in ccn6 morpholino-injected muscle: representation of five different experimental sets.
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