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Fig. 2. mTORC1 controls proximal tubular expansion. (A) Schematic of the mTORC1 signaling pathway indicating the three pharmacological inhibitors (rapamycin, Torin2, and LY294002) and their targets. (B) Schematic of the experimental layout. (C–J) pAkt and pS6 immunofluorescence analysis of stage 42 embryos either untreated or treated with 4 μM rapamycin, 1.2 nM Torin2, or 2 μM LY294002 from stage 34 onward. Tubular structures are outlined by yellow dotted lines. (K–O) 3G8 whole-mount immunohistochemistry and quantification of proximal tubular cells in untreated controls and embryos with inhibited mTORC1 signaling at stage 42. (P–R) Comparison of embryo length and skeletal muscle formation using the monoclonal antibody 12/101 in the presence or absence of 4 μM rapamycin at stage 42. Number of embryos analyzed and the SD are indicated in the individual bars. *P < 0.005. Image published in: Romaker D et al. (2014) Copyright © 2014. 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. Experiment + Assay Source Phenotypes and Disease Xla Wt + Torin 2 + NF42 (immunohistochemistry) fig.2.e Xla Wt + LY294002 + NF42 (immunohistochemistry) fig.2.f Xla Wt + Rapamycin + NF42 (immunohistochemistry) fig.2.h Anatomical Phenotype decreased ribosomal protein S6 kinase activity Xla Wt + Torin 2 + NF42 (immunohistochemistry) fig.2.i Anatomical Phenotype decreased ribosomal protein S6 kinase activity Xla Wt + LY294002 + NF42 (immunohistochemistry) fig.2.j Anatomical Phenotype decreased ribosomal protein S6 kinase activity Xla Wt + Rapamycin + NF42 (immunohistochemistry) fig.2.l Anatomical Phenotype abnormal proximal tubule morphology decreased size of the proximal tubule Xla Wt + Torin 2 + NF42 (immunohistochemistry) fig.2.m Anatomical Phenotype abnormal proximal tubule morphology decreased size of the proximal tubule Xla Wt + LY294002 + NF42 (immunohistochemistry) fig.2.n Anatomical Phenotype abnormal proximal tubule morphology decreased size of the proximal tubule Xla Wt + LY294002 + NF42 (immunohistochemistry) fig.2.o Anatomical Phenotype decreased size of the proximal tubule Xla Wt + Rapamycin + NF42 (immunohistochemistry) fig.2.o Anatomical Phenotype decreased size of the proximal tubule Xla Wt + Torin 2 + NF42 (immunohistochemistry) fig.2.o Anatomical Phenotype decreased size of the proximal tubule Larger Image Printer Friendly View

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