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. 2021 Sep 20;12:5536. doi: 10.1038/s41467-021-25551-1

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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 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 — a Scheme showing the expansion in a liquid−liquid interphase (LLI) followed by the differentiation at an air-liquid interphase (ALI) of normal human bronchial epithelial cells (NHBE). b Three-dimensional immunofluorescence analysis (z-stacks) of representative NHBE cells stained with antibodies specific for the indicated differentiation markers. Shown is one representative out of two biologically independent experiments. c NHBE cells were left untreated or infected with the indicated CoV (MOI = 3) and treated for up to three days with thapsigargin (0.1 or 1 µM). Supernatants were collected at five time points p.i. and virus titers determined by plaque assay. Data represent three biologically independent experiments using NHBE cells derived from two or three independent donors. Shown are means ± s.d. of technical duplicates. For HCoV-229E and MERS-CoV, cells from donor 1 were plated and differentiated a second time to generate an additional independent experiment (labeled donor 1.2). See Supplementary Fig. 5f for cell viability experiments.