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. 2020 Jul 17;3:388. doi: 10.1038/s42003-020-1090-2

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© The Author(s) 2020

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. 2 — a Changes in intracellular metabolite levels detected by GC-TOF with MTBSTFA (n = 4 biologically independent replicates), in Atg12^f/f and Atg12^KO MEFs in control media or following HBSS starvation for the indicated times. Fold change is relative to Atg12^f/f in control media. b–dAtg12^f/f and Atg12^KO MEFs in control media or following 2 h HBSS starvation were lysed and immunoblotted for markers of mTORC1 signaling (p-S6, p-4EBP1). b Representative immunoblot is shown. Relative protein levels of (c) p-S6 and (d) p-4EBP1 were quantified, normalized to loading control, and shown as a boxplot with dotplot overlay for each biological replicate. Statistical analysis by ANOVA with Tukey’s HSD post hoc test was performed and p values for Atg12^f/f samples compared with Atg12^KO samples in control media or HBSS are shown. e Representative immunoblot for markers of mTORC1 signaling (p-S6, p-4EBP1) in Atg12^f/f and Atg12^KO MEF protein lysate following a timecourse of HBSS starvation.