. Author manuscript; available in PMC: 2008 Jan 30.

Published in final edited form as: Dev Biol. 2005 Jun 15;282(2):550–570. doi: 10.1016/j.ydbio.2005.04.010

Table 2.

Effects of ectopic Hh pathway activation on hindbrain gene expression in yot mutants

yot allele (marker gene)	Injected RNA	Number of Embryos	Fraction of embryos with ectopic marker expression
			Wild-type (+/+ and +/−)^#	Mutant (−/−)^#
you-too^ty17(netrin1a expression at21 hpf)	none	137	0% (0/97)	0% (0/40)
	dnPKA^*	128	91% (84/92)	81% (29/36)
you-too^ty119(nk2.2 expression at21 hpf)	none	32	0% (20/20)	0% (12/12)
	dnPKA^*	60	88% (38/43)	0% (0/17)^@
you-too^ty119 (islet1-GFP expression at36 hpf)	none	27	0% (20/20)	0% (7/7)
	dnPKA^*	52	79% (30/38)	0% (0/14)^@

While dnPKA- injected yot mutants could be readily identified after in situ hybridization, roughly half of the embryos in each experiment was genotyped by PCR. Since there was perfect correlation between PCR genotyping and morphological identification, the data from embryos scored by the two methods were pooled.

Approximately 1 ng of dnPKA RNA (see Materials and methods) was injected per embryo, generating the Hh overexpression phenotype in 80–90% of injected wild-type embryos (Chandrasekhar et al., 1999).

Although no nk2.2-or GFP- expressing cells were found at ectopic locations in the yot^ty119 mutant hindbrain following dnPKA overexpression, expressing cells were found in the caudal hindbrain of several embryos, where nX neurons are normally found (see Figs. 3D, H). In addition, the number of GFP-expressing nVII neurons was markedly increased, and nk2.2 expression in ventral r4 was consistently upregulated in many of these embryos.