Xenbase Image ID: 175556

Fig. 2. Decreased function of Dsp in the embryo. (A) Schematic showing where the DspMO1 (red bar) is predicted to bind to dsp.L mRNA as well as the predicted spliced products in Dsp morphants and controls. Black arrow heads indicate the region of primer sequences used for RT-PCR analysis. RT-PCR of dsp showing there is an alternative splice product in the DspMO1 morphants. Immunofluorescence of desmoplakin (white) in DspMO1 and CMO morphants. (B) Schematic showing where the DspMO2 (red bar) is predicted to bind to dsp.L mRNA as well as the predicted spliced products in Dsp morphants and controls. Black arrow heads indicate the region of primer sequences used for RT-PCR analysis. RT-PCR of dsp showing there is an alternative splice product in the DspMO1 morphants. Immunofluorescence of desmoplakin (white) in DspMO2 and CMO morphants. (C) Schematic showing where the dspCrispr1 (red bars) is predicted to target the dsp.L and dsp.S genes. Black arrow heads indicate the region of primer sequences used for mutation analysis. Results of a T7 endonuclease assay are shown of dspCrispr mutants showing there is an alternative product indicative of mutation. Immunofluorescence of desmoplakin (white) in dspCrispr mutants and controls. (D) Schematic showing where the dspCrispr2 (red bars) is predicted to target the dsp.L and dsp.S genes. Black arrow heads indicate the region of primer sequences used for mutation analysis. Results of a T7 endonuclease assay are shown of dspCrispr mutants showing there is an alternative product indicative of mutation. Immunofluorescence of desmoplakin (white) in dspCrispr mutants and controls. E-I) Lateral view of control, Dsp morphants and mutant embryos at stage 28–30. Anterior is to the left. Arrows indicate regions where the epidermis is broken and arrowheads point to blister like structures. Scale bars = 450 μm. J-N) Later view of control, Dsp morphants and mutant embryos at stage 40–41. Anterior is to the left. Arrows indicate regions where the epidermis is broken and arrowheads point to blister like structures. Scale bars = 450 μm. O) Analysis of phenotypic frequency in i) DspMO1 morphants (D1), ii) DspMO2 morphants (D2), iii) dspCrispr1 mutants (Cr1), dspCrispr2 mutants (Cr2) compared to controls (C). Orange indicates percentage with the defect indicated at the top. Abbreviations: WT = wildtype uninjected sibling embryos, E = exon, IF = immunofluorescence. Image published in: Bharathan NK and Dickinson AJG (2019) Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V. Experiment + Assay Source Phenotypes and Disease Xla Wt + dsp MO + NF28-29/30 (whole-mount microscopy) Fig. 2 F Anatomical Phenotype abnormal epidermis morphology abnormal incomplete closing of the neural fold decreased size of the embryo increased pigmentation in the epidermis Disease skin disease Xla Wt + dsp MO + NF28-29/30 (whole-mount microscopy) Fig. 2 F Anatomical Phenotype abnormal epidermis morphology abnormal incomplete closing of the neural fold decreased size of the embryo increased pigmentation in the epidermis Disease skin disease Xla Wt + dsp CRISPR + NF28-29/30 (whole-mount microscopy) Fig. 2 H Anatomical Phenotype abnormal epidermis morphology decreased size of the embryo Xla Wt + dsp CRISPR2 + NF28-29/30 (whole-mount microscopy) Fig. 2 I Anatomical Phenotype abnormal epidermis morphology decreased size of the embryo Xla Wt + dsp MO + NF40-41 (whole-mount microscopy) Fig. 2 K Anatomical Phenotype abnormal craniofacial region morphology abnormal epidermis morphology abnormal eye morphology abnormal tail morphology decreased size of the embryo obsolete abnormally edematous heart Disease skin disease Xla Wt + dsp MO + NF40-41 (whole-mount microscopy) Fig. 2 L Anatomical Phenotype abnormal craniofacial region morphology abnormal epidermis morphology abnormal eye morphology abnormal tail morphology decreased size of the embryo obsolete abnormally edematous heart Disease skin disease Xla Wt + dsp CRISPR + NF40-41 (whole-mount microscopy) FIg. 2 M Anatomical Phenotype abnormal craniofacial region morphology abnormal epidermis morphology abnormal eye morphology abnormal tail morphology decreased size of the embryo obsolete abnormally edematous heart Xla Wt + dsp CRISPR2 + NF40-41 (whole-mount microscopy) Fig. 2 N Anatomical Phenotype abnormal craniofacial region morphology abnormal epidermis morphology abnormal eye morphology abnormal tail morphology decreased size of the embryo obsolete abnormally edematous heart Larger Image Printer Friendly View

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