Fig. 9. B56ε is required for the Hh pathway. (A) Whole mount in situ hybridization showing εmor (3.5 ng) injection blocked Shh (1 ng)-induced rx reduction (upper panels) and ptc-1 expression (lower panels) in whole embryos. Embryos were bilaterally injected at the 8-cell stage. (B) RT-PCR showing εmor (10 ng) injection blocked ptc-1 and foxA2 induced by shh (1 ng), smo-M2 (2.5 ng), and gli1 (0.5 ng) in neuralized animal caps. εmor and RNAs were injected sequentially at the 1-cell stage. Animal caps were dissected at stage 8/9 and harvested at stage 14. (C) RT-PCR showing εmor injection (10 ng) blocked ptc-1 expression induced by wild-type Gli1 (500 pg), but not by Gli-VP (50 pg and 100 pg). Embryos were injected and manipulated as described above. (D) Western blot showing overexpression of Shh inhibited the processing of overexpressed Myc-Gli3 (0.5 ng) in animal caps. The effect of Shh (1 ng) on Myc-Gli3 processing was not affected by εmor (10 ng) injection. Embryos were injected and manipulated as described above, except that caps were harvested at stage 12. (E) Co-IP results showing overexpression of Shh (1 ng) dissociated complex formation between FLAG-Cos2 (1 ng) and Myc-Gli1 (1 ng) in animal caps. Injection of εmor (10 ng) did not affect this complex dissociation induced by Shh. Embryos were injected and manipulated as described above. (F) Whole mount in situ hybridization showing the expression of rx in (from left to right) a control embryo, an embryo injected with εmor (3.5 ng), an embryo injected with Gli-VP (100 pg), and an embryo injected with Gli-VP and εmor. Embryos were bilaterally injected at the 8-cell stage.
Image published in: Rorick AM et al. (2007)
Copyright © 2007. Image reproduced with permission of the Publisher, Elsevier B. V.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| rax | rax-a, rax-b, rax1, rx, rx-1, rx1, rx1a, Rx2A, Xrax, Xrx1, Xrx1A | Xenopus | Throughout NF stage 18 | eye primordium |
| rax | rax-a, rax-b, rax1, rx, rx-1, rx1, rx1a, Rx2A, Xrax, Xrx1, Xrx1A | Xenopus | Throughout NF stage 20 | eye primordium |
| ptch1 | patched, patched1, Ptc-1, Ptc1, X-ptc1, XPtc1 | Xenopus | Throughout NF stage 18 | ectoderm neuroectoderm |
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