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. 2017 Aug 24;6(10):1472–1482. doi: 10.1242/bio.027359

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© 2017. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 5. — An increased number of NeuroD⁺ cells is unaccompanied by changes in neuroblast proliferation in Tbx1^Cre/−;Jag1^flox/+ and Tbx1^Cre/−;Jag1^flox/flox mutants at E9.5. (A) Bar graphs plotting the mean mitotic index per ear within the neuroblast population as defined by NeuroD expression. Mitotic index was calculated by dividing the total number of proliferating cells (pHH3⁺) that colocalized with NeuroD by the total number of NeuroD⁺ cells. Values were markedly lower in Tbx1^Cre/−;Jag1 ^flox/+ and Tbx1^Cre/−;Jag1 ^flox/flox OVs and the differences reached statistical significance. (B) Bar graphs plotting the mean total number of pHH3⁺ cells in the OV within NeuroD⁺ domain per OV. Values were statistically significantly lower in Tbx1^Cre/−;Jag1 ^flox/+ and Tbx1^Cre/−;Jag1 ^flox/flox OVs compared to Tbx1^Cre/− OVs. (C) Bar graphs plotting the mean total number of pHH3⁺ cells in the OV outside the NeuroD⁺ domain per OV. Values were statistically significantly lower in Tbx1^Cre/−;Jag1 ^flox/+ and Tbx1^Cre/−;Jag1 ^flox/flox OVs compared to Tbx1^Cre/− OVs. (D) Examples of the dual immunofluorescence experiment using antibodies to NeuroD and pHH3, used to calculate the above data. There are visibly fewer proliferating cells throughout the OV of Tbx1^Cre/−;Jag1 ^flox/+ embryos compared to Tbx1^Cre/ embryos. The VIIIth ganglion/CVG is markedly larger in Tbx1^Cre/−;Jag1 ^flox/+ mutants compared to Tbx1^Cre/− mutants. A distinct portion of the fused distal epibranchial ganglia (IX/X) is also outlined below. Note: three embryos and six OVs per genotype were used for each group. Embryos were stage-matched at 27ss. Data are mean±s.e.m. *P<0.05; **P<0.005.