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. 2018 May 17;8:7809. doi: 10.1038/s41598-018-24119-2

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© The Author(s) 2018

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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STED sub-diffraction image of hyper-tubulated peroxisomes. Peroxisome proliferation was induced by overexpression of PEX11β. (A) STED overview image of HeLa cell overexpressing PEX11β-myc fusion 24 hours after transfection, probed with a monoclonal anti-myc antibody and labeled with a secondary antibody conjugated to KK114 dye. (B,C) Blow up of hyper-tubulated PEX11β-myc structures. Arrow indicates a vesicle that may be budding of from the peroxisome. (D) Size (diameter) distribution of wild-type and hyper-tubulated peroxisomes. PEX11β-myc tubule’s diameter (probed with anti-Myc) was measured by FWHM of the Gaussian fit (N = 200 line scans from 70 peroxisomes from five cells). The mean diameter (d_mean) = 94.9 ± 21.8 nm for wild-type and (d_mean) = 91.8 nm ± 20.6 nm (±SD) for PEX11β-myc. For comparison, untransfected HeLa cells probed with anti-PEX14, quantified by distance between peaks of Gaussian fit membrane profiles (N = 74 profiles from 16 independent slices). Differences in the peroxisomal mean diameter were not significant (n.s) with p > 0.05. Scale bars 500 nm. (E) Histogram of peroxisome length showing PEX11β-myc hyper-tubulation >800 nm (N = 169 peroxisomes). Control: HeLa cells probed with the anti-PEX14 show no (N = 142 peroxisomes).