Expression of obscurin and obscurin-MLCK in developing zebrafish embryos. A: RT-PCR of total RNA from zebrafish embryos using primers from the 5' end of the obscurin/obscurin-MLCK gene. RNA samples were prepared from whole embryos 15, 22, 24, 36, 48, 60, and 72 hr post-fertilization (hpf) and from adult zebrafish hearts. Expression is first noted in embryos approximately 22 hpf with expression persisting in the adult heart. B: Immunolocalization of obscurin/obscurin-MLCK expression using the link7 and Ank antibodies in zebrafish embryos at 48 hpf. Note the expression of obscurin and obscurin-MLCK in the heart (∧) and skeletal muscle (arrows and inset, far right). Immunolocalization of myosin (MF20 antibody), which decorates the A bands of striated muscle, is shown for comparison. Note that, unlike myosin, both obscurin and obscurin-MLCK are also expressed in non-muscle tissues, including the central nervous system (*), at this stage in development.
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