Acquired CCN2 deficiency induces changes in the aortic gene expression pattern
RNA-sequencing was used to assess aortic transcriptomics changes and to provide insight into the molecular mechanisms involved in CCN2 actions in the vasculature and in aneurysm formation. As expected, a heat map two-way hierarchical clustering of genes and samples showed that CCN2-KO + Ang II mice clearly differed from the other sets of mice (Figure 6A ), but it also exhibited a different aortic gene expression pattern in the CCN2-KO mice compared to WT group (Figure 6B ). Although samples were analyzed as pools limiting the statistical power to detect differentially expressed genes (Rajkumar et al., 2015), we used them as hypothesis-generating experiments. To search for potential mechanisms involved in the vascular effects of CCN2 deletion, a GO enrichment analysis was done using the most deregulated genes obtained from the RNA-Seq studies in the WT vs . CCN2-KO comparative analysis (67 genes, Q-value < 0.5; fold-change >0.6) (Table 1 ). In this analysis, BP category results identified C21-steroid hormone biosynthetic process, cholesterol metabolic process, steroid biosynthetic and metabolic processes, or aldosterone biosynthetic process as some of the most deregulated terms in absence of CCN2. Regarding the CC level, extracellular region, extracellular space, secretory granule or mitochondrial crista were the standout terms, while Toll-like receptor 4 binding, scaffold protein binding, iron ion binding or oxidoreductase activity terms spotlight in the MF category (Table 2 ).