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. 2021 Mar 1;28(7):2194–2206. doi: 10.1038/s41418-021-00747-6

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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. 2 — A Representative confocal images of mtYFP fluorescence in thymocytes freshly isolated from WT;Lck-Cre⁺;mtYFP⁺ and Opa1^fl/fl;Lck-Cre⁺;mtYFP⁺ mice. Scale bar: 5 µm. B Representative electron micrographs of thymocytes freshly isolated from WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Scale bars: 2 µm or 500 nm as indicated. C Total thymocyte cellularity of WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Each symbol represents an individual mouse. Data are shown as flow scatter dot plots with mean ± SEM bars (N = 7/8 each group). D Representative flow cytometry of DN, DP, CD4⁺, CD8⁺ thymocyte subsets in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. E Frequencies of DN, DP, CD4⁺, CD8⁺ thymocyte subsets in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Each dot represents an individual mouse (N = 12 each group). Mean ± SEM are reported. F Cellularity of DN, DP, CD4⁺, CD8⁺ thymocyte subsets in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Each dot represents an individual mouse (N = 8 each group). Mean ± SEM are reported. G Representative flow cytometry of DN1–DN4 subsets (gated on lineage-negative DN thymocytes) in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. H Frequencies of DN1–DN4 thymocyte subsets (gated on lineage-negative DN thymocytes) in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Each dot represents an individual mouse (N = 7 each group). Mean ± SEM are reported. I Cellularity of DN1–DN4 thymocyte subsets (gated on lineage-negative DN thymocytes) in WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Each dot represents an individual mouse (N = 6/8 each group). Mean ± SEM are reported. J DN3 and DN4 thymocytes were sorted from WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice and OCR was measured at baseline and after exposure to oligomycin (Oligo), FCCP, and rotenone/antimycin (Rot/Ant). Data are shown as mean ± SEM (N = 4/group). K Basal OCR of DN3 and DN4 thymocytes isolated from WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. Data are shown as mean ± SEM (N = 4/group). L Principal component analysis of gene expression of DN3 and DN4 thymocytes isolated from WT;Lck-Cre⁺ and Opa1^fl/fl;Lck-Cre⁺ mice. M Gene expression of DN4 Opa1^fl/fl;Lck-Cre⁺ versus DN4 WT;Lck-Cre⁺ thymocytes analyzed by RNAseq. N Heatmap visualization of the 13 significantly differentially expressed genes between DN4 Opa1^fl/fl;Lck-Cre⁺ versus DN4 WT;Lck-Cre⁺ thymocytes. N = 3/group.