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. 2020 Jun 11;11:2967. doi: 10.1038/s41467-020-16754-z

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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. 5 — Placental cells were infected with ZIKV MOI of 1 for five days. a LD staining with the BODIPY 493/503 dye (green), anti-Flavivirus group antigen antibodies (Env) staining distinguishes ZIKV⁺ (red) from ZIKV⁻ bystander cells (asterisk). High-magnitude images of LD are shown in the right panels. Scale bar, 20 μm. High magnification image insets showing LDs in both ZIKV⁺ (Env⁺) and ZIKV⁻ (Env⁻) bystander. Scale bar, 5 μm. b Morphometric measurements of LD number and volume based on Z-stack projections of confocal images. Data from five different donors are shown as dot plot graphs. Each data point represents the average number of LDs per cell and average volume from mock (blue) and ZIKV-infected cells (red) or bystander neighboring cells (violet). Results are presented as mean values ± SEM and analyzed by one-way ANOVA, with Dunnett’s multiple comparison test. c Supernatants from mock- or ZIKV-infected primary placental cells were collected from five independent donors 5 dpi and UV-irradiated. Freshly isolated placental cells are challenged with the UV-irradiated supernatants for 3 days. Immunofluorescence staining of LDs with the BODIPY 493/503 dye (green), ZIKV Env protein (red) and nuclei (DAPI in blue). Scale bar, 50 μm. d LD number and volume analysis by Imaris software. Each data point represents the average number of LDs per cells and average volume from Mock- (blue) and ZIKV+ cells (red). Results are presented as mean values ± SEM and analyzed by two-tailed Student’s t test. Source data are provided as a Source Data file.