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. 2020 Jan 30;11:620. doi: 10.1038/s41467-020-14484-w

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© The Author(s) 2020

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. 3 — a Co-infection of Sf9 cells with recombinant baculoviruses (rBVs) containing the genes for TbIMPDH and Pex3-mCherry fusion protein 7 days p. i. The Pex3-marker, shown in a “fire” look-up-table for better visibility, labels peroxisomal membranes. The right panel shows that mCherry-fluorescence clearly surrounds TbIMPDH crystals. The white arrowhead points to a crystal vertical in the confocal plane, also surrounded by mCherry fluorescence. Middle panel: DIC image. Left panel: overlay. b Co-infection of rBVs containing the genes for TbIMPDH and cytoplasmatic EGFP 6 days p. i. Confocal imaging shows EGFP fluorescence within the crystal volume (white arrowhead), that could result from diffusion of EGFP molecules into channels within the TbIMPDH crystal within the cytosol. c Co-infection of rBVs containing the genes for TbIMPDH and peroxisomal EGFP-SKL 7 d. p. i. Left panel: confocal plane showing EGFP fluorescence within the crystal volume, but no enrichment around the crystal demonstrating the cytosolic localization of the crystal. Right panel: maximum projection from a complete Z-stack of the same cell showing many intense point-like structures but no enrichment of EGFP-SKL around the cytosolic crystal. d Co-infection of rBVs containing the genes for TbIMPDH and peroxisomal EGFP-SKL 8 days p. i. Right panel: EGFP fluorescence. Left panel: DIC. EGFP-SKL shows cytoplasmatic fluorescence, as well as localized enrichment, identified as peroxisomes. Plasma membrane disruption is visible after treatment with hypotonic buffer at the 0-s-timepoint. EGFP fluorescence is quickly lost from the cytoplasm and the crystal volume, whereas peroxisomes remain undisturbed (see Supplementary Movie 4).