Fig. 1. xDMO embryos have reduced xDnmt1 levels, are abnormal and mis-express genes. (A) Top panel: in vitro inhibition of xDnmt1 translation (black arrowhead) using xDMO (compare lanes 2 and 3). Bottom panel: in vivo inhibition of xDnmt1 translation in pre-MBT (stage 7-8) embryos (compare wild-type and xDMO extracts). Tubulin is used as a loading control. (B) Left panel: phenotypes of stage 15 embryos. Morphant xDMO embryos exhibit apoptotic lesions (arrowheads and enlargement) and lack neural folds (black arrow) compared with control stage 15 embryos. xDMO embryos contain fluorescein, unlike the control embryo (compare arrowed embryos). Right panel: comparison of percentage (n=100) of successfully neurulating embryos for wild type and xDMO. (C) xDMO embryos mis-express a range of transcripts. Wild-type and xDMO RNA was assayed by RT-PCR over a 10-fold dilution range (0.1, 0.3 and 1 µl cDNA for each sample indicated by the black triangles). H4 is a loading control. (D) In situ analysis reveals ectopic expression of the indicated xDMO targets throughout the animal pole (compare wild-type and xDMO panels). The maternally expressed gene xOct60 is not mis-expressed. Scale bars: 1 mm in B,D. Animal pole views are shown.