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Bissected chick caudal explant pairs treated '−'or '+' cyclopamine and then analysed by in situ hybridisation for Lfng or Gli1 mRNA expression.
(C ): Bissected chick caudal explant pairs treated '−' or '+' the inhibitors and then analysed by in situ hybridisation show expression of cLef1 is down-regulated after 3 hr treatment in the presence of XAV939 in the chick PSM.
Bissected chick caudal explant pair treated '−' or '+' 10μM roscovitine and 0.1 nM LY411575 and then analysed by in situ hybridisation for the Notch target gene cLfng reveals cLfng mRNA expression is delayed in the treated side as compared to the control after 3 hr treatment.
(E – J ): Bissected chick caudal explant pairs treated for 3 hr '−' or '+' the inhibitors and then subjected to fix and culture where the cultured side is cultured for 90 min (E, F ) or 2 hr (G – J ) and then analysed by in situ hybridisation for cLfng mRNA expression.
(A – D ): Bissected chick caudal explant pairs treated '−' or '+' the inhibitors and then analysed by in situ hybridisation for the Notch target gene cHairy2 reveals cHairy2 mRNA expression is delayed in the treated side as compared to the control after 3 hr treatment with XAV939 (A ), Roscovitine (B ), DRB (C ), PHA-767491 (D ).
The null hypothesis was rejected in the starred cases (p < 0.005) with the following p values XAV = 0.0211; Rosc = 0.0002; Pha = 0.0001; DRB = 0.0007 (D – H ) Bissected chick caudal explant pairs subjected to the fix and culture assay after 6 hr treatment in the presence of DMSO, XAV939, Roscovitine, PHA767491 or DRB.
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However, more recent observations suggested that, in the chick embryo, the caudal part of s0 epithelializes first [45], [68].
CdxA: chick; Chicken CHox-cad gene for caudal homologue, activator.
Conversely, misexpression of Hes1, an effector of Notch signaling, in the caudal hindbrain of the chick lowered the expression of Ascl1 in p3[5HT] progenitors to levels comparable to the expression of Ascl1 in spinal cord p3 progenitors (control p3[5HT] = 221.9 [mean] ± 20.9 [SD]; Hes1 electroporation p3[5HT] = 123.4 ± 8.7; control p3[V3] = 119.8 ± 22.1).
Nevertheless, DELTA-NOTC Hsignalling fulfils a different role in the growing caudal neural plate of the chick embryo.
As shown in Fig. 3D,E, electroporation of the caudal spinal cord of HH10 chick embryos with pCIG-Delta1 in scattered cells reproduced the endogenous expression pattern of Hes5.1.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com