Fig 1: SLC25A22 knockout abrogates secretion of CXCL1 and CXCL3 in vitro and in vivo.a RNA-seq of SLC25A22 knockout DLD1 cells and gene set enrichment analysis (GSEA) for the identification of common differentially regulated pathways in SLC-KO1 and SLC-KO2 cells (n = 4). b, c GSEA enrichment scores for differentially regulated gene sets unveiled the cytokine-cytokine receptor interaction signaling pathway as the top pathway depleted in SLC25A22 knockout cells. d Inflammatory Response and Autoimmunity PCR array showed that CXCL1, CXCL3 and IL1B were induced in ApcMin/+KrasG12D/+Villin-Cre mice tumors, but were down-regulated by SLC25A22 knockout (FC > 2). e qPCR validated that SLC25A22 knockout inhibited CXCL1/3 mRNA in DLD1, CT26 and Colo26 cells (n = 3). Each dot represents an independent sample. f Antibody array showed SLC25A22 knockout down-regulated cytokine secretion in DLD1 cells. g Densitometry showed CXCL1 and CXCL1/2/3 were top-down-regulated cytokines. h ELISA confirmed SLC25A22 knockout impaired CXCL1/3 secretion in DLD1 (72 h), CT26 (24 h) and Colo26 (24 h) (n = 3). Each dot represents an independent sample. i Detection of CXCL1/3 in serum and tumors of mice implanted with CT26 allografts (left, n = 5) and ApcMin/+KrasG12D/+ organoid allografts (right, n = 10) with or without SLC25A22. Each dot represents an independent mouse. j SLC25A22 mRNA correlates with CXCL1/2/3 mRNA in TCGA CRC (COADREAD) cohort (n = 677). Each dot represents an independent patient. Data are shown as mean ± SD (e, h, i). Two-tailed one-way ANOVA (e, h, i). Two-tailed Student’s t test analysis for two-group comparison i. Pearson correlation test j. Source data are provided as a Source Data file.
Fig 2: CD200R signaling regulates chemokine expression in TME(A) Differential gene expression analysis of the scRNA-seq data revealed significantly altered overall expression of chemokine genes in NB9464D tumors from CD200R−/− mice versus tumors from WT mice.(B) Violin plots for the expression of chemokine genes that show significant differences between tumors from WT and CD200R−/− mice. ∗∗∗P < 1e−100, ∗∗P < 1e−10 by Wilcoxon rank-sum test.(C and D) qPCR analyses for the expression of chemokine genes in NB9464D (C) and Yummer1.7 (D) tumors from WT and CD200R−/− mice. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by two-sided student t test.(E) ELISA assay for production of chemokines in NB9464D tumor lysates. ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by two-sided student t test.(F) Featured plot data of scRNA-seq suggests that CCL24 and CCL8 were mainly expressed by TAMs, while CCL3, CXCL2 and CXCL3 were mainly expressed by neutrophils, but also by TAMs.(G) Differential gene expression analysis of the scRNA-seq data revealed altered expression of chemokine genes in TAMs and Neutrophils.(H) qPCR was used to quantify expression of chemokine genes in sorted CD11b+Ly6G− and CD11b+Ly6G+ cells from NB9464D tumors from WT and CD200R−/− mice.
Supplier Page from Abcam for Mouse GRO gamma ELISA Kit (CXCL3)