Fig 1: Comparison of transcriptomic signatures and inflammatory phenotypes in astrocytes and BMEC-like cells as a function of co-culture and treatment with TNF.A PCA clustering plot of RNA-seq data in BMEC-like cell. B, C Volcano plot of DEGs (FDR < 0.1) in BMEC-like cells in monoculture versus co-culture, with or without TNF treatment. Red and blue points highlight genes associated with endothelial cell-cell junctions, internalization and trafficking, and angiogenesis. D, E GO analyses in the BMEC-like cells in monoculture versus co-culture. F PCA clustering plot of RNA-seq data in astrocytes. G, H Volcano plot of DEGs (FDR < 0.1) in astrocytes in monoculture versus co-culture, with or without TNF treatment. Red and blue points highlight genes associated with inflammation, chemokines, and immune responses. I, J GO analyses in the astrocytes in monoculture versus co-culture. K Heat map showing fold change of gene expression of maturity-related genes in co-cultured astrocytes compared to monocultured astrocytes. The gene set is based on ref. 47. L Heat map showing relative expression of genes associated with AD and inflammation across all astrocyte conditions. The gene sets are based on ref. 48 (left panel) and ref. 106 (right panel). The color of the heat map represents the log2(FPKM value) for each gene. M, N Representative immunofluorescent images of VCAM-1 expression in BMEC-like cells under different conditions (M) and select quantification (N). Data were acquired at day 14 according to the timing presented in Fig. 2J. Quantification was conducted using n = 3 biological replicates. The box shows the range between the 25th and 75th percentiles, the line within each box indicates the median, the outer lines extend to 1.5 times the interquartile range from the box, and each data point represents an individual image (3 images quantified per biological replicate). Statistical analyses were performed with a two-sided t test. O Representative western blot showing relative VCAM-1 and E-selectin expression in BMEC-like cells in monoculture or co-culture, with or without TNF treatment. Data were acquired at day 14 according to the timing presented in Fig. 2J. GAPDH was used as a loading control. Trends were confirmed across n = 3 biological replicates.