<i>dmd</i> mRNA and Dmd protein expression show differences in spatial expression.

<p>(a-d). Chromogenic <i>in situ</i> hybridization for <i>dmd</i> in wild type embryos at 18hpf (a,b) and 22hpf (c,d). (a,c) Wide field image of 18hpf and 22hpf embryos stained for <i>dmd</i> expression. (b,d) 3D projections of confocal z-stack of embryos in (a) and (c), respectively. Fluorescent signal of NBT/BCIP stain for <i>dmd</i> transcript reveals spatial confinement of transcript between somites. (e-p). Confocal image of <i>dmd</i>-<i>citrine</i> transcript (f,j,n) detected by <i>in situ</i> HCR and antibody staining for DMD-citrine protein (e,i,m) in <i>Gt(dmd-citrine)</i>^ct90a (e-l) and wildtype (m-p) embryos at 18hpf and 26hpf. Counter-stain with antibody to Ctnnb1 in blue (g,k,o). (h,l,p) Merged of image of (e-g, i-k, and m-o). Comparison of protein and transcript expression at 18hpf (e-h) and 26 hpf (i-l) in <i>Gt(dmd-citrine)</i>^ct90a embryos show that DMD protein localize exclusively to the myosepta while <i>dmd-citrine</i> mRNA is expressed in the cytoplasm at 18hpf and becomes more localized to the myosepta at 26hpf. Dmd-citrine expression in the nucleus can be seen as dots that appear similar in distribution between 18hpf and 26hpf. (m-p) DMD-citrine protein (m) and transcript (n) are not detected in wildtype embryos. Anti-Citrine antibody staining is in green, <i>citrine</i> mRNA HCR is in red and anti-Ctnnb1 antibody is in blue. Scale bars (a) 50μm (b,m) 20 μm.