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. 2024 Oct 30;16(12):3033–3056. doi: 10.1038/s44321-024-00157-4

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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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Figure EV2 — (A) Quantitative analysis of the absolute number of neutrophils (left) and macrophages (middle) within PDAC tumor tissues from control-vaccinated mice (v-CTRL) and ADAM12-vaccinated mice (v-A12); corresponding flow cytometry gating strategy of myeloid cells (right): neutrophils were identified as CD45⁺CD11b⁺Ly6G⁺ cells; monocytes were gated based on non-neutrophils (negative gating), and then identified as CD11b⁺Ly6C⁺ population. Then, monocyte macrophages were further defined with an F4/80⁺ signal. (n = 12 mice in v-CTRL, n = 12 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test). (B) Representative immunofluorescence staining images (left) and quantitative analysis of CD31-positive endothelial cells (right) on PDAC tumors from mice treated with either control vaccine (v-CTRL) or ADAM12 vaccine (v-A12). (n = 6 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm). (C) Representative immunofluorescence staining images (left) and quantitative analysis of pericyte coverage assessed by α-SMA/CD31 co-localization on PDAC tumors with either control vaccine (v-CTRL) or ADAM12 vaccine (v-A12) and representative images of α-SMA/CD31 immunostaining. (n = 6 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm). (D) Representative immunofluorescence staining images (left) and quantitative analysis of tumor hypoxia via GLUT1-positive cells on subcutaneous PDAC from v-CTRL mice and v-A12 mice together with representative images of GLUT1 immunostaining. (n = 6 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm). (E) Gene expression analysis of the KP2-specific gene CK19 in peripheral blood at endpoint (day 28 post tumor inoculation) from subcutaneous PDAC mouse model with control vaccine (v-CTRL) or ADAM12 vaccine (v-A12). (n = 5 mice in v-CTRL, n = 5 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test).