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. 2021 Jun 23;220(9):e202104069. doi: 10.1083/jcb.202104069

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© 2021 Ronchi et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S1. — Characterization of the behavior of mCherry fluorescence in organoids in resin block.(A and B) Fluorescence intensity measurements of H2B-mCherry in mammary gland organoids. Confocal stack of a representative sample in A. The integrated fluorescence intensity of 10 nuclei per confocal slice was measured, and the average (±SD) is plotted in B in relation to the distance from the block surface. The data are normalized to the average fluorescence intensity of the nuclei at the surface. (C and D) Bleaching curve of H2B-mCherry in a resin block. A FOV containing organoids was consecutively scanned 250 times, with the settings described in the Materials and methods section. Images are shown in C, and quantification of the integrated fluorescence intensity of nuclei from three independent organoids (±SD) is shown in D (red curve). Each experimental point in the chart is represented as a fraction of the fluorescence intensity in the first image. The bleaching experiment of H2B-mCherry in PFA-fixed hydrated Matrigel was conducted with the same settings used for resin-embedded organoids. An average of three independent organoids (±SD) is represented in gray.