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. 2022 Apr 11;24(4):483–496. doi: 10.1038/s41556-022-00869-0

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© The Author(s) 2022

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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Extended Data Fig. 1 — (a) RPE-1 cells treated with DMSO or MG132 were stained for CP110 and the indicated protein. Scale bar 2 μm. (b) The indicated cell lines were treated with MG132 and stained for pHSP27, CEP135 and PCM1. DNA was stained with DAPI. Scale bar 10 μm, inset 2 μm. (c) Quantification of the percentage cells forming an aggresome upon DMSO, MG132 and BZ treatment, as indicated. Data displayed as mean ± s.d., n = 3 independent experiments. ****p < 0.0001; ***p < 0.001. (d) RPE-1 cells treated with MG132 were stained for PCM1 and the indicated protein. Scale bar 2 μm. (e) Histogram showing the percentage of cells with greater than 4 foci of CEP135, SAS6, GT335, CETN2, CP110 and CEP97 following MG132 treatment. Data displayed as mean ± s.d., n = 3 independent experiments. ****p < 0.0001; ***p < 0.001; ns, not significant. p values: 0.218071 (CEP135), 0.058423 (SAS6), 0.160943 (GT335), 0.000219 (CETN2), 0.000055 (CP110), and 0.000150 (CEP97). (f) TEM images of centrioles from RPE-1 cells treated with DMSO or MG132. Scale bar 500 nm. (g) RPE-1 cells treated with DMSO or MG132 for 5 hours were stained for PCM1, pHSP27 and the indicated protein. Scale bar 2 μm. (h) Super resolution images of aggresomes in MG132 treated RPE-1 cells stained as indicated. Scale bar 1 μm. (i) Co-localization between the indicated protein pairs from individual z-planes of super resolution images of RPE-1 cells treated with MG132 displayed using Pearson correlation co-efficient. Boxes represent the median, upper and lower quartiles, whiskers represent 1.5x the interquartile range, with individual values from two independent experiments superimposed. For c,e: p determined by two-tailed unpaired Student’s t-test. Numerical data and p values are provided as source data.

Source data