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Supplemental Figure 2: Characterization of Dsp expression. A) RT-PCR of Dsp at embryonic stages as well positive controls in the adult skin and heart. RT+ = reverse transcriptase was added, RT- = reverse transcriptase was not added. Primer sequences available upon request. B-E''') Immunofluorescence of Dsp (green) in the epidermis from various embryonic stages. Dapi (blue) and beta-catenin (pink) labeling were used as a counter stain. F-F''') A 3D rendering of images as shown in E-E''' were created and then rotated 90 degrees. G-G''') st. 30 where only the inner epidermal layer of cells are shown. All scale bars = 40µm |
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Fig. 1. Desmosomes and desmoplakin in the epidermis of X. laevis embryos. A) Schematic showing a transverse section of the X. laevis embryonic epidermis based on actual histological sections. B) TEM images where black arrows indicate desmosomes located between outer epidermal cells. Scale bar = 1.8 μm. Inset shows higher magnification of a desmosome. Scale bar = 135 nm. C-D‴) Immunofluorescence of Dsp and beta-catenin in the epidermis. Dapi labeling of the nuclei was used as a counter stain. C-C‴) st. 19–20, in H arrows indicate regions where there is a less Dsp labeling and arrowheads indicate where there is enrichment of Dsp labeling. The same cells are indicated in panel C′. Scale bar = 27 μm. D-D‴) High magnification of a cell that appears to be apically emerging with high levels of Dsp and lower beta-catenin. Scale bar = 6 μm. D-D‴) High magnification of a cell that appears to be beginning to intercalate and has low levels of Dsp. Scale bar = 6 μm. Abbreviations = MCC = multiciliated cell, SVs = secretory vesicles, mito = mitochondria, TJ = tight junction, oe = outer epidermis, ie = inner epidermis. |
