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. 2022 Sep 8;54(9):1332–1347. doi: 10.1038/s12276-022-00802-3

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© The Author(s) 2022

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 — Images assessing the colocalization of MCT1 and mPCs in L6 cells, which show the localization of DAPI-positive nuclei (A), MCT1 (B), mPC1 (C), and MitoTracker-positive MR (D) in L6 cells. The merged images are shown in (E). Colocalization analysis of mPC1 (C) and mitochondria D showed a Pearson correlation coefficient (r²) value of 0.8. Colocalization analysis of MCT1 (B) and mPC1 (C) showed an r² of 0.3, largely because MCT1 occupies sarcolemmal, mitochondrial and peroxisomal compartments. A channel to represent the colocalization of MCT1 and mitochondria was created to image mMCT1; subsequent colocalization of mMCT1 with mPC1 resulted in an r² of 0.8 (F). White dots indicate the colocalization of mMCT1 and mPC1 as observed in ImageJ software. Whole images were contrast-enhanced in (A, B, C, D, and E). Similar results were observed for mPC2. Scale bar = 20 µm. It appears that both MCT1 and the putative mPC colocalized to the mitochondria (r² = 0.8). However, at the light microscopic level, it is impossible to know if the two proteins interact physically and functionally. Additionally, with the benefit of the Orbitrap LC/MS device, we could determine the fractional synthesis rates of mLOC and mPC proteins. From Brooks⁶.