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miR499 diminished the ossification of intermuscular bones in zebrafish. (A) Micro-CT imaging of whole-body skeletons, revealing reduced IB ossification area in miR499−/− zebrafish. en: epineural bone; ep: epipleural bone. Red dashed boxes indicate partial intermuscular bones (IBs); (B) Isolated IBs from zebrafish (scale bar = 500 μm); (C,D) Quantification of the number of epineural bones and epipleural bones. N = 5; (E,F) Quantification of IB ossification mean area at 3 mpf. N = 3. * p < 0.05, ** p < 0.01; (G) qRT-PCR analysis of bone-related gene expression in IB tissues separated from muscle. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01. ns: no significance.
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