IMAGE

Fig. 3

ID
ZDB-IMAGE-120724-20
Source
Figures for Flowers et al., 2012
Image
Figure Caption

Fig. 3 Notum 1a interacts with specific glypicans. (A) Inhibition of posterior body in notum1a-overexpressing embryos is enhanced by depletion of gpc3. Although co-injection of gpc3 MO increases the number of embryos with severe inhibition of posterior body (P=0.003), co-injection of gpc3 mRNA reduces the number of embryos displaying any posterior body impairment compared with notum1a-injected (P=0.05) and notum1a mRNA- and gpc3 MO-injected (P=0.002) embryos. The phenotypic classes are the same as depicted in Fig. 2A. (B) Notum 1a overexpression produces a faster migrating form of Gpc3-EGFP. A western blot of lysates from mGFP/gpc3-EGFP-injected and mGFP/gpc3-EGFP/notum1a-injected embryos probed with an anti-GFP antibody reveals that the gpc3-EGFP band migrates faster in lysates from notum1a-injected embryos. (C-F) Notum 1a overexpression does not mimic the Gpc4 loss-of-function phenotype. (C,D) Confocal images of the notochords of gpc4fr6/+ (C) and gpc4fr6/fr6 (D) Tg(Bactin:HRAS-EGFP)vu119 embryos at five somites. Although the cells of the notochords of gpc4fr6/+ embryos display normal mediolateral elongation and orientation, the cells are disorganized in the homozygous mutants. (E,F). Confocal images of three somite embryos injected with mCherry-HsHRAS RNA non-heat shocked and Tg(hs:notum1a) embryos heat shocked at 50% epiboly. Both control (E) and notum1a-overexpressing (F) embryos display normal cell elongation and orientation. (G) notum1a does not interact with Gpc4 or Wnt11. An incross of gpc4fr6/+ results in a Mendelian ratio of embryos displaying a mutant phenotype at 24 hpf. Injection of gpc4 mRNA efficiently rescues this phenotype. Co-injection of notum1a with gpc4 mRNA does not impair the rescue of the mutant phenotype (P=0.565). (H) Similarly, wnt11tz216/+ fish were incrossed to produce a Mendelian ratio of mutant embryos displaying some degree of cyclopia. This phenotype can be suppressed by injection of wnt11 mRNA, and co-injection of notum1a did not inhibit this effect (P=0.95).

Figure Data
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Development