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. 2024 Aug 28;15:7458. doi: 10.1038/s41467-024-51901-w

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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. 1 — a Experimental approach to measuring global translation rates in senescent cells. b–d O-propargyl-puromycin (OP-Puro) incorporation and protein synthesis rates in proliferating and senescent BJ-Ras-ER cells (b), IMR-90-Ras-ER cells (c), and TIG3-BRAF-ER cells (d). Data represent mean ± SD from biologically independent experiments (b, n = 8; c, d, n = 6). p-values were calculated using a two-tailed unpaired t test. ***P < 0.001. e Protein synthesis rates in proliferating and senescent cells treated with doxorubicin (DOXO) or camptothecin (CPT) for 7 days. A549, HCT116, HeLa, and MCF7 cells were used. Data represent mean ± SD from biologically independent experiments (n = 3). p-values were calculated using a two-tailed unpaired t test. ***P < 0.001. f OP-Puro incorporation assay in IMR-90 cells undergoing replicative senescence. PD, population doubling. Data represent mean ± SD from biologically independent experiments (n = 3). p-values were calculated using a two-tailed unpaired t test. ***P < 0.001. g Quantification of senescence-associated beta-galactosidase (SA-β-gal) expression in BJ-Ras-ER cells transduced with sgRNAs targeting the p53 gene or a control sequence. Data represent mean ± SD from biologically independent experiments (n = 5). p-values were calculated using a two-tailed unpaired t test. ***P < 0.001. h Quantification of bromodeoxyuridine (BrdU) incorporation in various CRISPR-Cas9-transduced BJ-Ras-ER cells. Data represent mean ± SD from biologically independent experiments (n = 3). p-values were calculated using a two-tailed unpaired t test. ***P < 0.001. i Quantification of protein synthesis rates based on OP-Puro incorporation in proliferating and senescent BJ-Ras-ER cells transduced with either a control sgRNA or sgRNAs targeting the p53 gene. Data represent mean ± SD from biologically independent experiments (n = 3). p-values were calculated using a two-tailed unpaired t test. NS, not significant; ***P < 0.001. Source data, including exact p-values, are provided as Source Data files.