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Fig. 1. PTK7 protein is required for posterior neural cell fate specification. (A) Left panel: sqRT-PCR to pools of ten early-neurula (st.12) embryos injected with 15 ng Meis3-MO at the one-cell stage. HoxD1 expression inhibition is as a positive control for Meis3-MO activity. His4 controls for RNA levels. A control PCR omitting reverse transcriptase (-RT) was performed on RNA isolated from control embryos in all experiments shown. Right panel: sqRT-PCR to pools of five neurula (st.19) embryos injected with 22 ng Meis3-MO at the one-cell stage. Krox20 expression inhibition is a positive control for Meis3-MO activity. EF1α controls for RNA levels. (B) sqRT-PCR to pools of six early-neurula (st.12.5) embryos injected with 10-20 ng PTK7-MO at the one-cell stage. (C) Left panel: embryos were injected into one blastomere at either the one- or two-cell stage with PTK7-MO (12 or 6 ng, respectively). Four posterior cell-type markers were analyzed by in situ hybridization. Neurula embryos (st.17), viewed dorsally, anterior is on top. Arrows and dashed line mark the dorsal midline; two-cell-stage embryos injected on right side. Krox20 expression (a) is normal in 100% of the control embryos (n=21) and perturbed (b,c) in 78% of the PTK7-morphant embryos (n=46). HoxB3 expression (d) is normal in all of the controls (n=23) and perturbed (e,f) in 98% of the PTK7 morphants (n=46). N-tub expression is normal (g) in all controls (n=23) and perturbed (h,i) in 95% of the morphants (n=44). Snail2 expression is normal (j) in all controls (n=20) and perturbed (k,l) in 87% of the morphants (n=46). Right panel: sqRT-PCR to pools of six neurula (st.19) embryos injected at the one-cell stage with PTK7-MO (10 or 20 ng). (D) Upper panel: sqRT-PCR to pools of eight neurula (st.17) embryos injected at the one-cell stage with PTK7-MO (12 ng) and RNA encoding the human PTK7 protein (100 pg). Lower panel: sqRT-PCR to pools of seven neurula (st.16) embryos injected at the one-cell stage with PTK7-MO (10 ng) and RNA encoding the Meis3 protein (1.1 ng). Image published in: Bin-Nun N et al. (2014) Copyright © 2014. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue egr2.L EGR-2, Krox-20, krox20, XKr20, XKrox-20 X. laevis Throughout NF stage 17 rhombomere R5 rhombomere R3 hoxb3.S hox2.7, hoxb-3, LOC108700804, LOC108702517 X. laevis Throughout NF stage 17 rhombomere R5 tubb2b.S N-Tub, n-tubulin, NBT, neural beta-tubulin, NST, ntubulin, Xn-tubulin X. laevis Throughout NF stage 17 Rohon-Beard neuron neuron snai2.L slug, snai2-a, snai2-b, Snail2, xSlu, xslug, XSnail2 X. laevis Throughout NF stage 17 neural crest cranial neural crest Image source: Published Larger Image Printer Friendly View

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