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. 2022 Apr 7;13:1895. doi: 10.1038/s41467-022-29335-z

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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. 4 — a Left panel, bright-field images of day 5 CFC derived from Blg-Cre;Brca1^f/f;Trp53^+/− and Blg-Cre;HMMR^Tg/Tg;Brca1^f/f;Trp53^+/− MECs transduced with EGFP-empty or EGFP-Cre lentivirus. Scale bar = 100 μm. Right panel, colony output and colony phenotype in day 5 CFC (inset, color-coded; mean ± standard error of the mean (s.e.m.) from n = 4 wells; n = 2 experiments; n = 100 cells/well). One-way ANOVA; ****p < 0.0001. b Left panels, lamin B1 (LMNB1) detection and/or 4ʹ,6-diamidino-2-phenylindole (DAPI) showing nuclei and nuclear lamina architecture in subconfluent MEC cultures. Scale bar = 40 μm. Right panels, quantification of nucleus size (μm²; mean ± standard deviation (s.d.) from n = 9 frames; n = 3 experiments; >125 cells/experiment). One-way ANOVA; ****p < 0.0001. c Frequency of micronucleus and nuclear blebbing in day 5 colonies (mean ± s.e.m. from n = 6 wells; n = 3 experiments; >125 cells/experiment). One-way ANOVA; ****p < 0.0001. d cGAS-positive micronuclei in Blg-Cre;HMMR^Tg/Tg;Brca1^f/f;Trp53^+/− MECs transduced with EGFP-Cre lentivirus. Scale bar = 10 μm. e Ratio (nucleus/cytoplasm) of p65 and p52 intensity in day 5 colonies (mean ± s.d. from n = 30 cells; n = 2 experiments). One-way ANOVA; ***p = 0.001. f Colony phenotype of Blg-Cre;HMMR^Tg/Tg;Brca1^f/f;Trp53^+/− EGFP-empty MECs treated with LPS during colony formation (mean ± s.e.m. from n = 6 wells; n = 3 experiments; 100 cells seeded per well). One-way ANOVA; ***p = 0.001. g Nucleus budding and micronuclei (LMNB1, left panel), and micronuclei (cGAS, middle panel) detection in HeLa cells overexpressing HMMR. Scale bar = 10 μm. Right panel, percentage of cells displaying nucleus budding or micronuclei (mean ± s.d. from n = 3 experiments with 105-242-253 (HeLa); 151-183-208 (Tet-On -dox); and 147-155-176 (Tet-On +dox) cells). One-way ANOVA; *p = 0.014. h Left panels, p52 nuclear/cytoplasm localization in analyzed cell settings. Scale bar = 30 μm. Right panel, p52 nuclear/cytoplasm ratio (mean ± s.d. from n = 3 experiments consisting of 150 (HeLa), 150 (-dox), and 150 (+dox) cells). One-way ANOVA; ****p < 0.0001.