Figure 6. Ackr4⁺ fibroblasts show elevated expression of multiple endothelial cell regulators.

(A) FACS gating strategy used to isolate Ackr4⁺ and Ackr4^- cells from among live single CD45^-GP38⁺CD31^- cells from the small intestine of Ackr4^gfp/+ (Ackr4^tm1Ccbl) mice. (B) Heatmap showing the relative signal intensities of the 165 transcripts that were differentially expressed between the Ackr4⁺ and Ackr4^- cells (Fold Change ≥2; adjusted p value <0.05; FDR 0.1). (C) Volcano plot showing differences between Ackr4⁺ and Ackr4^- cells in their expression of all genes analyzed, against the statistical significance of those expression differences. Genes shown in red have a Fold Change ≥2 and an adjusted p value of <0.1: those on the right hand side are more highly expressed in Ackr4⁺ cells; those on the left are more highly expressed in Ackr4^- cells. Ackr4 and genes analyzed in panel E are highlighted. Significance was calculated using a modified t-test, and adjusted using a Benjamini Hochberg multiple testing correction. (D) The biological processes most significantly upregulated in Ackr4⁺ cells compared to Ackr4^- cells, as determined using DAVID Bioinformatics Resources. (E) Relative expression of eight genes in Ackr4⁺ and Ackr4^- cells, as determined by QPCR analysis of cDNA from cells FACS-sorted from among live single CD45^-GP38⁺CD31^- cells from the small intestine. Gene names are indicated at the top of each graph. Data points from individual mice are shown, along with means (±1SD). Mean expression in Ackr4^- cells is set to 1. *p< 0.05, **p < 0.01, ***p< 0.001; Student’s t test.