Rnasol

Mice were mice treated with telmisartan for 3 weeks, their thoracic aortas placed in 400ul of RNAsol (Sigma Aldrich) and homogenized (OMNI International). RNA was extracted after overnight precipitation in isopropanol and resuspended in RNAse free water + 1:20 superase. RNA sample integrity was tested as previously described using the Agilent Bioanalyzer 2100^{25 (link)}. All computational analyses of RNA-seq data were performed in R (v 3.6.1). Genes with less than 2 counts per million (CPMs) in at least two samples were removed. Batch correction was applied using Combat-seq under the sva package (v. 3.35.2) with genotype and treatment specified as biological covariates²⁶ . Batch-adjusted data were processed and analyzed using DESeq2 (v 1.24.0)²⁷ with a design formula accounting for genotype, treatment, and their interaction including normalization, principal component analysis (PCA), and Wald test differential expression analysis. Heatmaps represented regularized log-transformed gene expression normalized by Z-score. Pathway enrichment analysis (Gene Ontology GO) was performed using clusterProfiler (v 3.12.0)^{28 (link)}. Log-transformed p-values and the number of genes mapped to each annotation are depicted on the dot plot.