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. 2021 Sep 17;12:5507. doi: 10.1038/s41467-021-25540-4

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© The Author(s) 2021

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. 5 — a Cell viability of Nalm-6 cells from monoculture (black, n = 8) vs 3T3-L1 adipocyte coculture (blue, n = 8) in the presence of the indicated chemotherapeutic agents or mitosis-independent stressors (no FBS and hydrogen peroxide). Data are presented relative to vehicle controls. b Cell viability of Nalm-6 cells after +48 h recovery from 3T3-L1 adipocyte coculture (blue, n = 4) or from monoculture (black, n = 4) after exposure to mitosis-dependant chemotherapy or mitosis-independent stressors (no FBS and hydrogen peroxide). Data are presented relative to vehicle controls. c Cell viability of Nalm-6 monocultured (black, n = 3) or 3T3-L1 adipocyte-cocultured Nalm-6 cells (blue, n = 3) in the presence of GCN2ib treatment (5 µg/mL) and the indicated cellular stressors. d Schematic for pulsed SILAC-based proteomic analysis. Nalm-6 cells cocultured with either 3T3-L1 adipocytes or preadipocytes were pulsed at +24 h and at 48 h with either ‘heavy’ or ‘medium’ isotope-labelled amino acids. Following a 4 h incubation, Nalm-6 cells were separated from their microenvironment for protein extraction. Lysates were mixed in equal amounts (between heavily and medium labelled samples at each time point), digested, fractionated and analysed using mass spectrometry.e Scatter plot showing the normalized log₂ ratio of proteins detected following pulsed SILAC-based proteomic analysis in Nalm-6 cells cocultured with 3T3-L1 adipocytes (Adipo) relative to preadipocytes (preadipo) at +24 h vs 48 h from one experiment. Blue Arrows describe the direction of change in adipocyte-specific coculture. Data are presented as mean values ± SEM values of independent experiments (n = 2 in (a) with four replicates, n = 1 with four replicates in (b, c). Statistical significance was assessed using 2-sided unpaired t tests (a, b for the single-agent treatments), Mann–Whitney U test for combination chemotherapy treatment (b) One-way ANOVA followed by Tukey’s test for multiple comparisons c. **p < 0.01, ***p < 0.001, ****p < 0.0001.