| |
Figure S3. Effect of the modulation of Notch signaling on the development of NC and surrounding tissues. (A) Design of the Delta1 MO. The Delta1 translation start site is indicated in red (a). Delta1 MO inhibits the translation of both Delta1 pseudoalleles (b). In vitro transcription-translation reactions were performed by using the Sp6 Rabbit Reticulocyte Lysate System (Promega) in the presence of 35S methionine. Translation products were analysed by SDS-PAGE/autoradiography.
(B) Whole-mount in situ analysis of embryos injected with Delta1 mRNA (1 ng), DeltaStu mRNA (1 ng) or Delta1 MO (7,5 ng) as indicated. The antisense probe used are indicated on the left. Note that Delta1 overexpression represses N-tubulin and increases Snail2 and Sox10. Conversely, N-tubulin is increased in Delta1Stu mRNA or Delta1 MO injected embryos (4 ng) and Snail2 and Sox10 is reduced. With the exception of Msx1 which is slightly reduced by the injection of DeltaStu mRNA or Delta1 MO, the other tested markers were unaffected by modulation of Delta1. Frequency of embryos with the indicated phenotype: a, 89% ectopic, n = 19; b, 57% reduced, n = 21; c, 60% reduced, n = 16; d, 45% increased, n = 20; e, 46% reduced n = 22; f, 67% reduced n = 18; g, 100% normal, n = 17; h, 82% normal n = 22; i, 72% normal, n = 22; j, 75% normal, n = 32; k, 77% normal, n = 22; l, 80% normal, n = 20 ; m, 100% normal, n = 26; n, 35% slightly reduced, n = 20; o, 40% slightly reduced, n = 24; p, 100% normal, n = 20; q, 100% normal, n = 18 ; r, 91% normal, n = 11; s, 100% normal, n = 10; t, 100% normal, n = 14 ; u, 100% normal, n = 16; v, 63% decreased, n = 19 ; w, 73% increased, n = 11; x, 38% increased, n = 8. Neurula stage embryos are viewed from the dorsal side, anterior down, with the injected side oriented to the right. Control and injected sides of tailbud embryos are shown. LacZ was used as a lineage tracer. |
