Aortic samples of inducible vascular smooth muscle specific KO of YAP/TAZ

YAP and TAZ are transcriptional co-activators and downstream effectors of the Hippo pathway, which play crucial roles in organ size control and cancer pathogenesis. Genetic deletion of YAP/TAZ has revealed their critical importance for embryonic development of the heart, vasculature and gastrointestinal mesenchyme. The aim of this study was to determine the long term role of YAP/TAZ in adult vascular smooth muscle in vivo. We used the novel Itga8-CreERT2 mouse for deletion of YAP/TAZ (i8-Y/T-KO). Overall design: RNA was isolated from control and i8-Y/T-KO thoracic, two weeks and eight weeks after the first tamoxifen injection, using RNeasy Mini Kit (Qiagen, 85600) and a QIAcube (Qiagen) workstation as per the manufacturer's recommendations. RNA concentration and purity was measured in a NanoDrop 2000c (Thermo scientific). RNA from Ctrl (n=4 at 2w and n=4 at 8w) and i8-YT-KO (n=4 at 2w and n=4 at 8w) thoracic aorta was sent to Novogene Europe (Cambridge, UK) for 150 bp paired-end sequencing on an Illumina platform. After cDNA library construction, sequencing, and filtering, reads were mapped to the Mus musculus reference genome (GRCm38/mm10) using Hisat2. Read numbers mapping to each gene were obtained using FeatureCounts and converted to Fragments Per Kilobase of transcript per Million mapped reads (FPKM). Differential gene expression analysis was done using DESeq2 in R. Enrichment analysis was performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) using clusterProfiler.