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. 2022 Jan 21;79(2):85. doi: 10.1007/s00018-022-04128-1

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — USP4 regulates the speed of co-localization of PDGFRβ with early endosomes. a, b Induction of USP4 delays co-localization of PDGFRβ with early endosomes. USP4 expression was induced in BJhTERT-USP4 tet-inducible cell line versus noninduced control cells. Representative images of co-localization (in yellow) of PDGFRβ (red) with early endosomal marker EEA1 (green) at indicated times of stimulation with PDGF-BB (a). Co-localization of PDGFRβ and EEA1 was quantified in 3 independent experiments and is presented as Pearson coefficient of correlation of distribution of signals for each analysed channel in 15–35 cells per condition. *p < 0.05 (b). c–e CRISPR-Cas9-mediated knockout of USP4 leads to faster co-localization of PDGFRβ with early endosomes. Co-localization of PDGFRβ and EEA1 marker was quantified and presented as in panel b for BJ-CRISPR-Cas9 knockout cells versus BJhTERT control cells (c) and U2OS CRISP-USP4 knockout cells versus U2OS control cells (d). e Representative images of co-localization (in yellow) of PDGFRβ (red) with early endosomal marker (green) at indicated times of stimulation with PDGF-BB of the experiments quantified in panel c are presented. f Co-localization between PDGFRβ and late endosomal marker (Rab7) is presented for BJhTERT CRISP-USP4 knockout cells versus BJhTERT control cells