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. 2022 Mar 3;41:81. doi: 10.1186/s13046-022-02300-w

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — CCL5-dependent fibroblast recruitment is mediated by SLC25A24 in fibroblasts. a Heatmap of the differentially expressed genes in the transcriptome sequencing of CCD-18Co before and after 40 ng/ml CCL5 stimulation for 24 h. Data, n = 3. b Relative expression of SLC25A24 mRNA in CCD-18Co and human primary normal colorectal fibroblasts (NF1, NF2, NF3, NF4, NF5, NF6, NF7, NF8, NF9) before and after 40 ng/ml CCL5 stimulation for 24 h. Data, mean ± SD; n = 3. c Representative α-SMA and SLC25A24 IF images at the invasive front of human CRC tissue without (the upper panel) or with (the lower panel) tumor budding trend. Data, n = 10. Scale bar, 100 μm. d Representative SLC25A24 IHC staining images of mouse subcutaneous tumors with the injection of SW620/shCtrl or SW620/shCCL5. Data, n = 3. Scale bar, 50 μm. e Model diagram of recruitment assay of 40 ng/ml CCL5 to SLC25A24 knockdown fibroblasts. f Immunoblots for SLC25A24 protein expression of CCD-18Co/siCtrl and CCD-18Co/siSLC25A24, NF4/siCtrl and NF4/siSLC25A24, NF7/siCtrl and NF7/siSLC25A24. g Quantification of cell numbers of CCD-18Co/siCtrl and CCD-18Co/siSLC25A24, NF4/siCtrl and NF4/siSLC25A24, NF7/siCtrl and NF7/siSLC25A24 recruited by 40 ng/ml CCL5. Data, mean ± SD; n = 5. h The differentially expressed genes in the transcriptome sequencing were analyzed by KEGG enrichment analysis. i Immunoblots for SLC25A24, PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR protein expression in CCD-18Co before and after different concentration of CCL5 stimulation for 24 h (the left panel) and those proteins in CCD-18Co/siCtrl and CCD-18Co/siSLC25A24 after 40 ng/ml CCL5 stimulation for 24 h (the right panel). SLC, SLC25A24; p-PI3K, phosphorylated PI3K; p-Akt, phosphorylated Akt; p-mTOR, phosphorylated mTOR. ns, no significance; *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed Student’s t test (b, g)