Fig. 7: Dysregulated CXCL16⁻S1PR1 signaling disrupts angiogenesis in GE.

a,b, A wheel plot (a) and heat map (b) from NicheNet analysis reveal ligand–receptor pairs between CD45⁺ immune cells and CD31⁺ endothelial cells dysregulated in GMH cases. The color intensity in the heat map indicates interaction potential (b). c, Violin plots show upregulated CXCL16 expression in most CD45⁺ subtypes in GMH cases. The data from a–c are from two independent biological samples in control and GMH cases. MG, microglia. WM, white matter. d,e, Images (d) and quantification (e) of Cxcl16⁺ and S100A9⁺ cells in GEs of control and GMH cases. Arrows in d indicate CXCL16⁺ and S100A9⁺ cells. f,g, Images (f) and quantification (g) of ELANE/CXCL16-treated microvessels show disorganization of VE–cadherin and actin and increased vascular permeability. h,i, Images (h) and quantification (i) of HUVEC branching morphogenesis treated with VEGF or VEGFand CXCL16. j, Violin plots show the expression of CXCR6 and S1PR1 in endothelial cells from control or GMH samples. k, Confocal and IMARIS 3D images of IBA1⁺ cells interacting with IB4⁺ vasculature in E12.5 control and Cdh5⁻Cre;S1pr1^fl/fl mice. White boxes indicate areas in the confocal images where enlarged IMARIS 3D images are captured. l, IBA1⁺ cell density and the percentage of IBA1⁺ cells touching blood vessels in the GE and cortical plate (CP) of E12.5 control and Cdh5⁻Cre;S1pr1^fl/fl mice. m, TEM shows myeloid cells inside the vascular lumen and in the brain parenchyma in the MGE of E12.5 control and Cdh5⁻Cre;S1pr1^fl/fl mice. The subdivisions (i–iv) show further magnification of TEM images. n,o, Confocal images and quantification show increased volume of CD68⁺ vesicles in IBA1⁺ cells in the GE of Cdh5⁻Cre;S1pr1^fl/fl mice (n) compared with the age-matched control (o). The dotted white lines define boundaries of the GE and CP where IBA1⁺ cells are quantified. p,q, Confocal images and quantification show increased density of CXCL16⁺ cells in the GE of Cdh5⁻Cre;S1pr1^fl/fl mice (p) compared with age-matched control (q). Statistics in c, e, g, i, l, o and q use a two-tailed, unpaired Student’s t-test, and the data represent the mean ± standard error of the mean. n.s., not significant. n indicates the number of independent biological samples used for quantification.