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. 2024 Aug 7;633(8029):433–441. doi: 10.1038/s41586-024-07803-4

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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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Extended Data Fig. 3 — a. Representative microphotographs of CCA H&E staining. Area between two black dotted lines represents intima. Green dotted lines represent necrotic cores. Necrotic core area was quantified as the percentage of total intima area (ANOVA, n = 8–10 per group). b. Representative images of smooth muscle actin (SMA) immunofluorescence staining. SMA area was quantified as the percentage of total intima area (ANOVA, n = 8–10 per group). c. Representative microphotographs of Picro-Sirius red staining. Collagen area was quantified as the percentage of total intima area (ANOVA, n = 8–10 per group). d. Representative images of Picro-Sirus red stained CCA sections with the according grid for fibrous cap (FC) thickness quantification. FC thickness was quantified at the locations were the FC crossed the applied grid (ANOVA, n = 8–10 per group). e. Representative microphotographs of CD68 immunofluorescence staining. Images were segmented by thresholding to convert fluorescence signal into a binary image. CD68 area was quantified as the percentage of total intima area (ANOVA, n = 8–10 per group). f. Left: Principal component analysis (PCA) using CCA plaque vulnerability readouts from sham, stroke and stroke mice with detected secondary lesion found in a. to e. (n = 8–10 per group) Right: Contribution of the Plaque vulnerability readouts in principal component 1 to 5 (PC1 to PC5), weighted for their relative quality of representation in the PCA. g. Arrows indicate a disrupted fibrous cap in lesion. The pie charts illustrate the proportion of mice with ruptured CCA plaques 1 w after sham or stroke surgery (n = 11 or 25 mice per group)(all scale bars = 50 µm).