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. 2021 Jan 18;11:1666. doi: 10.1038/s41598-021-81076-z

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Gene expression and mitochondrial status in LSK cells from control and Atg5^f/f;Vav mice. (a) Flow cytometric analysis of GFP-LC3 in LSK cells from control (GFP-LC3;Atg5^f/w;Vav and GFP-LC3;Atg5^f/w) and GFP-LC3;Atg5^f/f;Vav mice at P7 and 3W. Representative data (two control and two Atg5^f/f;Vav) from two independent experiments are shown (P7, n = 5–7; 3W, n = 3–5). (b) GSEA enrichment score curve for control (Atg5^w/w;Vav) and Atg5^f/f;Vav LSK cells analysed for the gene set associated with the mitochondrial protein complex. Heatmap of representative gene expression in the gene set is shown at the right. NES, normalised enrichment score; FDR, false discovery rate. (c) Flow cytometric analysis of MitoTracker green, MitoSox Red, TMRM, and DCFDA staining in LSK cells from control and Atg5^f/f;Vav mice at P7 and 3 weeks. Representative data from at least three independent experiments are shown (n = 3–5). (d) Fold change of representative Nrf2-target gene expression levels upregulated in Atg5^f/f;Vav LSK cells compared with control LSK cells from normalised microarray data. (e) mRNA expression levels of Nqo1 and Gstm1 in LSK cells from control, Atg5^f/f;Vav, p62^f/f;Vav, and Atg5^f/f;p62^f/f;Vav mice at 7 weeks. Data are the mean ± SD. n = 3–6. (f) Frequency of dead cells (7-AAD-positive cells) in LSK population from control (Atg5^f/w;Vav) and Atg5^f/f;Vav mice at P7 and 3W. Data are the mean ± SD (n = 4–5).