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Fig. S2. Hormone-inducible Notch constructs produce the same pronephric phenotypes as non-inducible Notch constructs.X. laevis embryos were injected into a V2 blastomere with hormone-inducible notch-ICD-hGR mRNA and su(H)DBM-hGR mRNA, left to develop to stage 18 then incubated in culture medium containing dexamethasone and DMSO (induced), or just DMSO alone (non-induced). Embryos were then left to develop to stage 34 when whole-mount in situ hybridisation for nephrin and slc5a2 was performed. (A, B) Embryos injected with notch-ICD-hGR mRNA that was activated at stage 18 showed a reduction in slc5a2 expression (A), but in non-induced embryos, slc5a2 expression was not affected (B). (C, D) Similarly embryos injected with su(H)DBM-hGR mRNA that was activated at stage 18 had reduced slc5a2 expression (C), but when not incubated in dexamethasone, these embryos displayed no pronephric phenotype (D). (E, F) Embryos injected with notch-ICD-hGR mRNA that was subsequently activated at stage 18 had ectopic nephrin expression (E), but when not activated no glomus abnormalities were observed (F). When embryos injected with su(H)DBM-hGR mRNA had their message activated at stage 18, nephrin expression was reduced. (G) Non-activated su(H)DBM-hGR mRNA had no effect on nephrin expression. Inducible Notch constructs produce similar phenotypes to non-inducible constructs and therefore the non-inducible constructs used in this work have specific effects on early pronephrogenesis. Asterisk denotes injected side. Image published in: Naylor RW and Jones EA (2009) Copyright © 2009. Image reproduced with permission of the publisher 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

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