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. 2019 Sep 30;27(4):1398–1414. doi: 10.1038/s41418-019-0426-2

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2019

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 — Time-lapse microscopy of PCD in a sA5-YFP embryo and during cranial neural tube closure. a Tracking of PCD events in an E8.75 sA5-YFP embryo upon induction of massive cell death by medium starvation (one frame every 5 min over 15 h, z-stack with 4 µm slices; images depicted are from slice #17, boxed images are close-ups from regions with arrows). After 2 h, sA5-YFP⁺ rounded cells (white arrows) appeared. b Fluorescence intensity from a over time. c, d Time-lapse microscopy of the brain of an E8 sA5-YFP embryo: the area marked with a white box in a is magnified in the subsequent pictures in d. After 18 min, the first rounded sA5-YFP⁺ cells (white arrows) appeared; notice that these cells stayed (dancing like) in this area without being engulfed for about 6 h. e Quantitation of sA5-YFP fluorescence signals from d over time. sA5-YFP secreted human Annexin V-yellow fluorescent protein, TL transmission light, and Afl autofluorescence. Bars are 100 μm (a, c) and 40 μm (d)