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. 2021 Jan 21;11:542950. doi: 10.3389/fphys.2020.542950

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Copyright © 2021 Haskins, Bhuvanendran, Anselmi, Gams, Kanholm, Kocher, LoTempio, Krohmaly, Sohai, Stearrett, Bonner, Tuchman, Morizono, Jaiswal and Caldovic.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Use of super-resolution microscopy to determine in situ nanoscale organization of urea cycle enzymes. (A) Schematic representing the possible distribution of urea cycle enzymes in the mitochondria. (B) Confocal images showing localization of CPS1 and the mitochondrial protein TOM20 labeled with Alexa Fluor 532 and Alexa Fluor 647, respectively. (C) Confocal and (D,F,G) gSTED images showing individual mitochondria in mouse hepatocytes immuno-stained for (C,D) CPS1, (F) OTC, (G) NAGS, (H) OTC (Alexa Fluor 647) and NAGS (Alexa Fluor 532), (J) CPS1 (Alexa Fluor 647) and NAGS (Alexa Fluor 532). Single labeling in images (C,D,F,G) was done with Alexa Fluor 647. (E,I,K) Normalized pixel intensity plot for the dotted lines marked in the corresponding confocal or STED images.