Panel A shows a schematic of the experimental setup for RNA-sequencing on resting bone marrow-derived macrophages (BMDMs) from wild-type (WT), VRAC knockout (KO), and Cre control, as well as WT and KO BMDMs incubated in hypo-osmotic media for 4 hours. Panel B is a scatter plot of principal component analysis (PCA) of gene-wise read counts, with axes labeled PC1 (54.69 percent) and PC2 (13.08 percent), showing distinct clusters for different conditions. Panels C, D, and E are volcano plots of differentially regulated genes (DEGs) in resting WT macrophages vs WT in hypo-osmotic media, resting KO macrophages vs KO in hypo-osmotic media, and WT macrophages in hypo-osmotic media vs KO macrophages in hypo-osmotic media, respectively. The axes are labeled Log2FC and minus log10(padj), with significant genes highlighted. Panel F is an enrichment map of cellular processes differentially regulated between WT and WT treated with hypo-osmotic media, with nodes representing genes and edges representing interactions.
Changes in macrophage cell volume induce a large transcriptomic response that alters diverse cellular processes. (A and B) RNA-seq was performed on resting BMDMs from WT, VRAC KO, or CX3CR1 Cre–expressing controls (Cre), and WT and KO BMDMs incubated in hypo-osmotic media (50% vol/vol H2O in DMEM) for 4 h (n = 3/condition). (B) Principal component analysis of the gene-wise read counts from A. (C–E) Volcano plots of differentially regulated genes (DEGs) in: resting WT macrophages vs WT in hypo-osmotic media (C), resting KO macrophages vs KO in hypo-osmotic media (D), WT macrophages in hypo-osmotic media vs KO macrophages in hypo-osmotic media (E). (F) Enrichment map of cellular processes differentially regulated between WT and WT treated with hypo-osmotic media.