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. 2019 Apr 26;10:1955. doi: 10.1038/s41467-019-09943-y

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — Hph-dependent amino-acid and oxygen sensing via Tor is HIF-1a/Sima-independent. a Growth response to dietary amino-acid levels (0.1×, 0.3×, and standard 1× dietary yeast concentrations) of wild types (w¹¹¹⁸) and Hph and sima mutants under normoxia (21% O₂) and hypoxia (5% O₂). Note that sima mutants are significantly larger than wild types on 1× yeast food in hypoxia, whereas their sizes are not significantly different in normoxia (p = 0.26), further indicating that Sima is required for hypoxia-induced growth suppression. n = 8–71. b Fat-body staining against phosphorylated ribosomal protein S6 (anti-pS6), a downstream indicator of Tor activity, indicates that the Tor pathway is suppressed by 16 h amino acid starvation (1× sugar-only medium), hypoxia (5% O₂), or Hph mutation. sima mutants exhibit Tor-pathway activation under normoxia, and thus Sima is not required for Tor activity. Tor activity decreases under hypoxia and with amino-acid deprivation, indicating that Sima activity is also not required for Tor suppression. Combined treatments of amino-acid starvation and hypoxia do not further suppress Tor. Representative images of pS6 stain are shown. n = 8–30. c Sizes of fat-body lipid droplets examined by CARS microscopy in wild types (w¹¹¹⁸) and Hph and sima mutants with 16 h hypoxia (5% O₂) or amino acid starvation (1× sugar-only medium). Lipid-droplet size increases under hypoxia and protein starvation in wild types and sima mutants, but not Hph mutants. Representative CARS images are shown. n = 10–18. d Hph activity reflects amino acid availability. Fat-body imaging of the GFP::ODD Hph-activity reporter indicates that GFP::ODD degradation is reduced under 4 h amino acid starvation (1x sugar food), suggesting that Hph activity is blocked by lack of amino acids. n = 20. Statistics: Student’s t-test for pairwise comparisons and one-way ANOVA with Dunnett’s for multiple-comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control. Error bars indicate SEM. Underlying data are provided in the Source Data file