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Fig. 5. Phenotypical analysis of ADAM19 KD at the gastrula stage. (A) Embryos were microinjected with 10 ng of either the MO19 or a control MO at the one-cell stage and allowed to develop until gastrula stage (stages 12 and 13). Whole mount in situ hybridization were performed using the markers Brachyury, Sox2 (Xbra Sox2, stage 12) and Chordin and Goosecoid (Chd and Gsc, stage 13). For all embryos, the dorsal view is presented with the anterior up. The red arrowheads point to the notochord. The number of embryos and the frequency of the phenotype depicted in the photographs are indicated in the text. (B) Quantitative real-time PCR was performed on embryos injected with either MO19 alone or together with ADAM19 Rescue mRNA (MO19 + A19R). The relative abundance of Chordin (Chd), Goosecoid (Gsc) and Brachyury (XBra) gene expression was measured by the 2(-) method using either embryos injected with the control MO or non-injected and normalized to α-tubulin. There was no effect of the control MO to gene expression. The asterisks represent statistical significance at P < 0.05 using a Student t test. There is a small decrease in Xbra (16%, not significant) and a robust increase in both Chd (187%) and Gsc (230%) which are rescued by the injection of the ADAM19 mRNA. |
