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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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Fig. 2 — a, RPE-1 cells treated with DMSO, MG132, cycloheximide (CHX), or CHX plus MG132 were stained as indicated. b, The area occupied by pHSP27 in cells treated as in a. n = 493 (DMSO), 630 (MG132), 459 (CHX) and 437 (CHX + MG132) aggresomes examined over 2 independent experiments. ****P < 0.0001. c, Cells treated with MG132 with or without pretreatment with CHX were stained as indicated. d, Intensity maps of PCM1 distribution relative to the centrosome in cells treated as in a. The percentage of PCM1 signal residing in the defined inner region is indicated. AU, arbitrary units. e, RPE-1 cells treated with DMSO or MG132 were treated concurrently with ACY-1215 or ACY-738 and stained as indicated. f, The area occupied by pHSP27 in cells treated as in e. n = 586 (DMSO), 566 (MG132), 470 (ACY-1215), 504 (ACY-1215 + MG132), 470 (ACY-738) and 395 (ACY-738 + MG132) aggresomes examined over 2 independent experiments. ****P < 0.0001. g, RPE-1 cells treated with MG132 with or without ACY-1215 or ACY-738 were stained as indicated. h, PCM1 distribution relative to the centrosome in cells treated as in e. i, RPE-1 cells were pretreated with nocodazole (Noc) or taxol followed by DMSO or MG132 and stained as indicated. j, The area occupied by pHSP27 in cells treated as in i. n = 554 (DMSO), 499 (MG132), 413 (Noc), 422 (Noc + MG132), 510 (taxol) and 485 (taxol + MG132) aggresomes examined over 2 independent experiments. ****P < 0.0001. k, PCM1 distribution relative to the centrosome in cells treated as in i. l, RPE-1 cells treated with MG132 and nocodazole were stained as indicated. For b, f and j, boxes represent the median, upper and lower quartiles, whiskers represent 1.5× the interquartile range, with individual values superimposed. Data were compared using Kruskal–Wallis analysis of variance (ANOVA) test, and post-hoc Dunn multiple comparison test was performed to calculate P values. Scale bars, 2 μm (a,c,e,g,i; inset of l) or 10 μm (l). Numerical data and P values are provided as source data. AU, arbitrary units.

Source data