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. 2022 May 13;13:867989. doi: 10.3389/fgene.2022.867989

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Copyright © 2022 De Rosa, Fasano, Zerillo, Valente, Izzo, Mollo, Amodio, Polishchuk, Polishchuk, Melone, Criscuolo, Conti, Nitsch, Remondelli, Pierantoni and Paladino.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The homeostasis of early endosomes is altered in DS fetal fibroblasts. (A) Human fetal fibroblasts from euploid (EU-HFFs) and DS (DS-HFFs) fetuses were stained with early endosome antigen 1 (EEA1) antibody detected with Alexa-546-conjugated secondary antibodies. Serial confocal sections were collected from the top to the bottom of the cells. Representative images showing that the early endosomes are enlarged in trisomic cells when compared to euploid cells. For each condition, the 3D reconstruction and corresponding intensity map are shown. Scale bars, 8 μm. The mean fluorescence intensity (arbitrary unit, a.u.) of EEA1-positive structures is higher in DS-HFFs than in EU-HFFs. The bars show the relative mean value ± SD of three independent experiments, n ≥ 50 cells. (B) Representative immunoblotting of EEA1 in DS-HFFs and EU-HFFs. Tubulin was used as the loading control. The molecular weight of protein markers is indicated. Densitometric analysis of three different experiments, performed in three euploid and in four trisomic cultures, is shown. Error bars, mean ± SD. ***p < 0.001, Student’s t-test.