Figure 4
Combining IH with epinephrine significantly alters epinephrine-induced changes in gene expression. HAECs were exposed to 60 cycles of IH (5%-20% O2) in the presence of 10 µM epinephrine and vehicle control. We then isolated RNA from 3 biologic replicates and performed RNA-sequencing. (a) Multidimensional scaling plot and (b) Volcano plots of DEGs in HAECs treated with normoxia + epinephrine and IH + epinephrine. (c) Heat map displaying the expression level of top epinephrine-upregulated genes in HAECs treated with epinephrine or vehicle control under normoxic and IH conditions. “Pretty heatmaps R package pheatmap package” from Z-score normalized expression values. https://cran.r-project.org/web/packages/pheatmap/pheatmap.pdf (d) Using qPCR, we measured mRNA expression of MT1G measured by qPCR in HAECs treated and untreated with 10 µm epinephrine under IH and normoxia (mean ± SD, n = 6, biologic replicates). *p < 0.05. (e) Multidimensional scaling plot of DEGs in HAECs treated with IH in the presence or absence of epinephrine (10 µM). (f) Volcano plot of DEGs in HAECs treated with IH in the presence or absence of epinephrine. Metallothionein-1G (MT1G) is highly differentially upregulated under IH supplemented with 10 µM epinephrine. (g) We also measured mRNA expression of epinephrine-upregulated genes, JUNB, EFNA1, IRF1, and NR4A1 in HAECs treated with epinephrine or vehicle control under normoxic and IH conditions (mean ± SD, n = 6, biologic replicates). *p < 0.05.
