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. 2024 Oct 19;15(10):761. doi: 10.1038/s41419-024-07116-4

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© The Author(s) 2024

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 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. 3 — A Left panel: Western blot analyzes the expression of LAP2α in U2OS (left) and SAOS2 (right) cells transiently transfected with control siRNA or LAP2α siRNA for 24 h and then re-expressing doxycycline (DOX)-inducible RNAi-resistant LAP2α for 48 h. Right panel: Quantification of the LAP2α relative protein level in comparison to GAPDH and to siNC was calculated by ImageJ software. Error bars represent the mean ± SEM of four independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. ns, not significant or p ≥ 0.05; *p < 0.05; **p < 0.01. B Representative images show the colocalization of telomere (green, detected by FISH) and PML (red, detected by immunofluorescence) to measure ALT-associated PML bodies (APBs) formation upon LAP2α knockdown in U2OS cells (left panel). Quantification of the average number of APBs in the U2OS cells (right). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. ***p < 0.001. C The APBs formation upon LAP2α knockdown in SAOS2 cells (left panel). Quantification of the average number of APBs (right panel). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. ***p < 0.001. D C-circle assay in ALT-positive cell lines U2OS cells (left panel). Quantification of the average amount of C-circle (right panel). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. **p < 0.01; ***p < 0.001. E C-circle assay in ALT-positive cell lines SAOS2 cells (left panel). Quantification of the average amount of C-circle (right panel). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. **p < 0.01; ***p < 0.001. F The colocalization of telomere (red, FISH) and 53BP1 (green, IF) to measure dsDNA breaks at telomere generated by the CRISPR/Cas9 system (left panel). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. ***p < 0.001. Quantification of the percentage of cells with TIFs (right panel). G C-circle assay in Hela cells (left panel). Quantification of the average amount of C-circle (right panel). Error bars represent the mean ± SEM of three independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. **p < 0.01; ***p < 0.001.