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. 2021 Jun 25;11:13317. doi: 10.1038/s41598-021-91747-6

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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 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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Subcellular localization of APRT1 and 2 in the BSF T. brucei cells. (A) Western blot analysis of the BSF cells overexpressing V5-tagged APRT1 and 2 which were treated with digitonin to obtain cytosolic and organellar fractions. The purified fractions were analysed with the following antibodies: anti-V5, anti-enolase (cytosol), anti-hexokinase (organellar fraction, glycosomes), and anti-mt hsp70 (organellar fraction, mitochondria). The protein marker is indicated on the left. (B) Immunofluorescence microscopy of the tetracycline induced (IND) V5-tagged APRT1 and 2. The tagged proteins were visualized using a monoclonal V5-antibody and an anti-mouse secondary antibody conjugated with fluorescein isothiocyanate (FITC). The MitoTracker Red was used to visualize mitochondria, while the DAPI was used to stain the DNA content [nucleus (n) and kinetoplast (k)] of the cell. WCL whole cell lysate, ORG organellar fraction, CYT cytosolic fraction.