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. 2023 Jan 9;25(1):108–119. doi: 10.1038/s41556-022-01042-3

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© The Author(s) 2023

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. 9 — (a) Representative immunoblot for LAP1 expression levels in primary melanoma WM983A cells stably expressing GFP-NLS and LAP1C-mRuby3, LAP1B-mRuby3 (LAP1 M122A) or LAP1-mRuby3, related to Fig. 5a–d. (b, c) Percentage of less metastatic A375P cells stably expressing GFP-NLS, A375P stably expressing GFP-NLS and LAP1-mRuby3, LAP1B-mRuby3 or LAP1C-mRuby3 that translocated their nuclei (b) and displayed nuclear envelope blebs (c) after a second round of transwell migration through 5-μm pores. n = 374, 316, 344 and 439 cells, respectively. (d, e) Percentage of less metastatic melanoma A375P cells stably expressing GFP-NLS and LAP1B-mRuby3, LAP1B^Δ1-72-mRuby3, LAP1B^ΔCBR-mRuby3 or LAP1B^R563G-mRuby3 that translocated their nuclei (d) and displayed nuclear envelope blebs (e) after a second round of transwell migration through 5-μm pores. n = 344, 412, 273, and 317 cells, respectively. (f, g) Percentage of primary melanoma WM983A cells stably expressing GFP-NLS and LAP1B-mRuby3 or LAP1B^Δ1-72-mRuby3, or siRNA resistant LAP1B-mRuby3 (LAP1B^R-mRuby3), LAP1B^Δ1-72-mRuby3 (LAP1B^Δ1-72R-mRuby3) or LAP1B^ΔCBR-mRuby3 that translocated their nuclei (f) and displayed nuclear envelope blebs (g) after a second round of transwell migration through 5-μm pores upon treatment with LAP1 siRNA#1. n = 322, 269, 305, 357, 368, 363, 400, 369, 372 and 375 cells, respectively. Note, OnTarget LAP1 siRNA#1 targets a sequence present within the CBR of LAP1. (h, i) Percentage of WM983A cells stably expressing GFP-NLS and LAP1C-mRuby3 or LAP1C^R441G-mRuby3 that translocated their nuclei (h) and displayed nuclear envelope blebs (i) after a second round of transwell migration through 5-μm pores. n = 418 and 388 cells, respectively. Graphs show the mean and error bars represent S.E.M. from N = 3 independent experiments. P-values calculated by one-way ANOVA (b-g) and two-tailed t-test (h, i; n.s.), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Unprocessed western blots, numerical data and exact p-values are available in the Source Data.

Source data