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Figure S1. tctp mRNA expression in the developing frog embryo. (A and B) In situ hybridization detection of tctp and pax6 mRNA expression on coronal sections of stage 43 embryos. The boxed areas, which mark the lateral surface of the mesencephalon through where retinal ganglion cell axons pass en route to the optic tectum, their synaptic target, are shown at a higher amplification in (A’) and (B’). (C) Continues Fig. 1D,E: Quantitative in situ hybridization detection of tctp mRNA expression in the RGC axonal and growth cone compartments was performed using stage 32 eye explants grown in vitro for 24 hours. Here, an additional control is shown – the hybridization signal obtained with labelled tctp mRNA was eliminated by competition using an 5:1 excess amount of unlabelled tctp antisense oligonucleotide probes. Quantification of mean fluorescence intensity is also presented (mean ± SEM; *** P < 0.0001, one-way ANOVA and Bonferroni); ‘sense’ fluorescence signal was used for normalization purposes. (D) Micrographs of the laser-capture microdissection procedure. (E) First 25 nucleotides of the Xenopus laevis tctp 5’UTR. As noted, the 5’UTR of tctp transcripts starts with a canonical 5’-terminal oligopyrimidine (TOP) motif, a typical feature of transcripts selectively regulated at the translational level by mTORC1. Scale bars: 100 μm in (A) and (B), 5 μm in (C), 200 μm in (D). Image published in: Roque CG et al. (2016) © 2016. This image is reproduced with permission of the journal 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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