RACK1 & neural tube closure, Wehner et al, 2011

1. RACK1 & neural tube closure, Wehner et al, 2011 Sklar,Podoly,Soreq (also McCahill, A.)

2. RACK1 & neural tube closure, Wehner et al, 2011 Boudeau, J.etal 2006 PTK7

3. RACK1 & neural tube closure, Wehner et al, 2011

4. RACK1 & neural tube closure, Wehner et al, 2011

5. RACK1 & neural tube closure, Wehner et al, 2011 Frog gastrul/neurul: http://www.youtube.com/watch?v=qisrNX3QjUg&NR=1

7. RACK1 & neural tube closure, Wehner et al, 2011

8. RACK1 & neural tube closure, Wehner et al, 2011

9. Wnt (wg) signaling pathway

10. RACK1 & neural tube closure, Wehner et al, 2011

11. RACK1 & neural tube closure, Wehner et al, 2011

12. RACK1 & neural tube closure, Wehner et al, 2011

13. RACK1 & neural tube closure, Wehner et al, 2011 RACK1 is a novel interaction partner of PTK7 that is required for neural tube closure DEV056291 Supplementary Material Files in this Data Supplement: •Supplemental Figure S1 - Fig. S1. The RACK1 MO1 causes neural tube closure defects if targeted to the neural ectoderm. (A) Injections were targeted to the posterior neural ectoderm by injecting into one dorsal animal blastomere at the eight-cell stage. For linage tracing, MOs were co-injected with GFP RNA. (B) Bar chart summarizing the percentage of neural tube closure defects of two independent experiments using increasing concentrations of the RACK1 MO. As a control, 20 ng control MO was injected. (C) Control MO-injected embryos show a closed neural tube, whereas RACK1 MO1-injected embryos show defects in neural tube closure. The upper panel shows an overlay of the GFP fluorescence and the bright-field image, the lower panel just the bright-field image. •Supplemental Figure S2 - Fig. S2. Neural tube closure defects of embryos injected with different combinations of PTK7, RACK1 and PKCδ1 MOs. Embryos were injected at the one-cell stage and neural tube defects were determined at neurula stage 19-20. (A) Additive effects are observed if 10 ng RACK1 MO was co-injected with 10 ng PTK7 MO. (B) Interestingly, co-injections of 10 ng PKCδ1 MO and 10 ng PTK7 MO support a synergistic interaction of PKCδ1 with PTK7. (C) Additive effects are also seen if 10 ng RACK1 MO was injected with 10 ng PKCδ1 MO. •Supplemental Figure S3 - Fig. S3. RACK1 is not required for FZ7-mediated DSH recruitment or hyperphosphorylation of DSH. (A) FZ7 recruits DSH to the plasma membrane in ectodermal explants injected with 100 pg DSH-GFP RNA, 100 pg FZ7-myc RNA and 20 ng control MO. (B) Co-injection of 10 ng RACK1 MO1 and 10 ng RACK1 MO2 does not affect the FZ7-mediated membrane localization of DSH. (C) Bar chart summarizing the percentage of cells with membrane-localized FZ7 and DSH. (D) Western blot, using anti-myc antibodies, of animal cap lysates injected with combinations of 100 pg DSH-myc RNA, 100 pg FZ7 RNA, 10 ng RACK1 MO1, 10 ng RACK1 MO2 or 20 ng control MO. Hyperphosphorylated DSH (P-dsh) is visible as a higher molecular weight band.

14. RACK1 & neural tube closure, Wehner et al, 2011 Sklar,Podoly,Soreq