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. 2023 Oct 24;14:6761. doi: 10.1038/s41467-023-42400-5

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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 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. 4 — a Baseline ccfDNA concentrations of healthy individuals (n = 19) and patients with recurrent or relapsed unfavorable CUP (n = 34). Each dot represents a single healthy individual or CUP patient, respectively; orange dots indicate CUP patients who radiologically respond to ICI treatment. The box contains the 25th to 75th percentiles of the dataset, a vertical line denotes the median value. The whiskers go from first or third quartile to the minimum or maximum values, respectively. An exact p-value (p = 0.0003) was calculated using a two-tailed Mann–Whitney U test. Of note, the y-axis is in log-scale for display purpose only. b Kaplan-Meier estimate of OS, stratified according to high (n = 12) or low (n = 17) ccfDNA based on a ccfDNA concentration cut-off of 5.2 ng/ml plasma. c The efficiency of the combined targeted/sWGS sequencing strategy in detecting ctDNA in blood samples of CUP patients. d Scatter plot of baseline ccfDNA concentration versus baseline ctDNA content of unfavorable CUP patients (n = 34). Each dot represents a single patient; orange dots indicate CUP patients who radiologically respond to ICI treatment. Correlation analyses were performed based on Spearman’s correlation coefficient; a regression line was fitted. Of note, the log-log axes are for display purpose only. e Kaplan-Meier estimates of PFS and OS, according to high (n = 14) or low (n = 15) ctDNA based on a ctDNA level cut-off of 37.4 hGE/ml plasma. In (b, e), crosses denote censored observations, and for each time interval the number of patients at risk are indicated below the plots. Comparisons are made using a two-sided log-rank test, Cox proportional hazard regression modeling was used to calculate hazard ratio. 95% CI 95% confidence interval, HR hazard ratio, IQR interquartile range, hGE haploid genome equivalent. Source data are provided as a Source Data file.