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. 2024 Dec 2;14:29913. doi: 10.1038/s41598-024-80677-8

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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 — Colorectal cancer cells exhibit low or absent Enpp1 expression and require Enpp1 transfection to degrade cGAMP. (a) RNA expression of (i) Enpp1, (ii) cGas, and (iii) Sting1 in a panel of breast, colorectal, and lung adenocarcinoma cell lines present in DepMap portal. Each symbol represents one cell line, with expression as Log2 transcripts per million. (b) Analysis of scRNASeq of a panel of murine tumors highlighting Enpp1 and Sting1 expression in (i) Cancer cells and (ii) stromal cells in MC38 tumors. Scale shows degree of expression by color and the circle size shows the percentage of each cell type that expresses the gene. (c) Analysis of scRNASeq of CD45 + cells in MC38 tumors gated on myeloid cells. (i) TSNE plot showing 6 clustered myeloid populations from MC38 tumors. (ii) Enpp1 expression (red) in clustered populations. Iii) identification of clusters based on key gene expression. (d) (i) Flow cytometry for Enpp1 versus isotype control in murine CT26 and MC38 colorectal carcinoma cell lines, and bone marrow-derived macrophages (BMM0). (ii) Summary of MFI of (i) across replicates. (iii) Recovery of spiked cGAMP from MC38 cells (NT), MC38 cells stably transfected with human Enpp1 (huEnpp1), human Enpp1 T256A mutant (*huEnpp1), or murine Enpp1 (muEnpp1). Key: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.