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DVDY_22313_sm_SuppFig1.tif5078KSupporting Figure 1: Expression of Amer genes during postimplantation mouse embryonic development. A: Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of Amer genes in embryonic day (E) 7.5–E10.5 mouse embryos. Expression of the Hprt gene was used as a control of mRNA levels. The expression of the three Amer genes at E10.5 was used as a positive control for the assay. B: Whole-mount in situ hybridization analysis of Amer genes at E9.0. Wtx/Amer1 was diffusely expressed along the neuraxis, with a higher expression level in the anterior part (white arrowhead). This anterior-posterior gradient of expression was more evident for Amer2. Note the strong staining located in the anterior embryonic territories and prospective forebrain (black arrowhead). The dotted line indicates the approximate level of a gelatin section after whole-mount staining (lower inset). No Amer3 expression could be detected at this stage. C: Whole-mount in situ hybridization of Amer3 at E9.5 showing high levels of expression in the anterior part of the developing neural tube and cranial ganglia. The white asterisk is used to indicate trapping of stain within the neural tube. (a,b) Gelatin sections of the stained embryos at a level shown in the schematic drawing. White arrowheads are used to indicate regionalization of Amer3 expression in the neural epithelium. Fv, fourth ventricle, H, heart; Hb, hindbrain; Me, mesencephalon; Nt, neural tube; S, somites; Sc, spinal cord; Tc, telencephalon; VII/VIIIg, facio-acoustic ganglion; IXg, petrosal ganglion; Xg, nodose ganglion.

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