Fig. S1

(A) Wild-type and MOdmd injected zebrafish embryos at 2 dpf, showing drastic reduction of Dystrophin on embryos injected with morpholino cocktail compared with wild type expression. No secondary developmental defects were observed. (B) Control quantitative PCR with specific primers for crebbp of mouse origin performed on 2 dpf control embryos (non-injected) and embryos injected at one-cell stage with 10⁷ (low), 5×10⁷ (medium) or 10⁸ (high) molecules of mRFP-CBP. Note that: i) endogenous zebrafish crebbp transcripts are not detected in control non-injected embryos; ii) the number of transcripts detected at 2 dpf increases linearly with the increasing number of transcripts injected at one cell stage. (C) Representative confocal sections of 2 dpf wild type embryos injected at one-cell stage with 10-fold doses of RFP-tagged CBP: 10⁷ (low) and 10⁸ (high) RNA molecules per embryo. Embryos were immunostained for exogenous CBP (αRFP; red), Dystrophin (green), and nuclei counterstained with DAPI (blue). Note the distribution of exogenous CBP, accumulated in the nuclei of a small subset of muscle cells when a lower dose is injected (B, red) compared with more widespread expression when a higher dose is injected (C, red). (D) Representative confocal tiled section of a 2 dpf embryo, co-injected at one-cell stage with MOdmd plus 10⁷ RNA molecules (low dose) of RFP-tagged CBP expression plasmid. Panels show immunostaining for Dystrophin (green), exogenous CBP (αRFP; red) and nuclei counterstained with DAPI (blue). Note that despite Dystrophin expression is undetectable the muscles are not damaged, even if only a small subset of total nuclei express exogenous CBP. (E) Control for z-crebbpa and z-crebbpb qPCR primers specificity: neither detects the exogenous CBP of mouse origin. PCR was performed on template pCS2+mRFP-CBP cDNA at concentrations: 100 ng/µl (lines 2-4), 10 ng/µl (lines 5-7) and 1 ng/µl (lines 8-10), and primers: m-crebbp (lines 2, 5, 8), z-crebbpa (lines 3, 6, 9) and z-crebbpb (lines 4, 7, 10). A 100-bp ladder (Biolabs) was used on lines 1 and 11.