XB-IMG-80375
Xenbase Image ID: 80375
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Figure 6. P2Y11 was expressed in the notochord, and was required for normal CE.
(A) Expression pattern of P2Y11, from the dorsal view (upper), and the cross section of the whole embryo at stage 11 and of the dorsal region at stage 17 (lower). (B) In vitro transcription/translation system showing the effect of P2Y11-MO on the transcription of P2Y11 mRNA. P2Y11-MO specifically inhibited the transcription of UTR-including P2Y11 mRNA (UTR-P2Y11-vns), constructed to generate a fusion protein with eYFP (vns). The effect of P2Y11-MO was restored by coexpression of a rescue construct (res-P2Y11-vns). P2Y11 protein was detected by western blotting with an anti-GFP antibody. Vns protein was detected as a loading control. (C) The proportion of phenotypes in control or P2Y11 morpholino (30 ng) -injected embryos at stage 28. Full length of P2Y11 mRNA lacking MO-targeting site (res- P2Y11 mRNA) rescued the phenotype with dose dependency. (D) Inhibition of nodal-expressing animal cap and Keller Explant elongation by the P2Y11-morpholino. The inhibition was rescued by res- P2Y11 mRNA overexpression. (E) The average length () of nodal-expressing animal caps. The elongation of animal cap was attenuated by P2Y11-morpholino and rescued by res- P2Y11 mRNA overexpression. (** p<0.01) (F) The average of the neck length () of Keller Explant with control or P2Y11 morpholino. The P2Y11-morpholino-injected explants were shorter and thicker than control morpholino-injected explants, and res- P2Y11 mRNA rescued its inhibition. (** p<0.01) (H) RT-PCR analysis of the nodal-overexpressing animal cap. Image published in: Shindo A et al. (2010) Shindo et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |