Fig. 5

Chronic and acute oxidative stress modulate iBMEC gene expression. a Principal component analysis of RNA collected from iBMECs seeded on Transwells in the presence of ROCK inhibitor one hour after acute exposure (d0), four days after acute exposure (d4), and seven days after chronic exposure (d7), and their respective vehicle controls. Each sequencing sample is pooled across two technical replicates for n = 4 biological replicates for each condition and time-point. b–d Volcano plots of differentially expressed genes in oxidatively stressed samples compared to their respective vehicle controls at days 0, 4, and 7 after exposure. e–f Heatmaps of adjusted p-value of selected pathways of MSigDB Hallmark Pathway and GO Biological Processes terms, respectively, from genes with log₂(fold-change) > 3. g Fold-change comparison of key pathways from GSEA indicating up- or down-regulation at day 4 after acute exposure vs. hour 1 after acute exposure. Pathways highlighted here were selected by sorting all resultant pathways by combined score (top 10 for MSigDB Hallmark and top 50 for GO Biological Processes). Pathways where one time-point had a combined score of zero were excluded. h Comparison of enriched genes, specific to key pathways for functional analysis, showing unique enrichment in acute or chronic oxidative stress profiles