The transcription factor Scleraxis differentially regulates gene expression in tenocytes isolated at different developmental stages

Purpose:- RNA sequencing was performed on adult and fetal equine tenocytes expressing a specific short hairpin RNA against scleraxis (shSCX) versus a non-target (NT) scrambled control Methods:- mRNA profiles of four independent biological lines of adult and fetal NT and shSCX expressing tenocytes were generated by RNA sequencing, using Illumina NovaSeq6000. The sequence reads that passed quality filters were analyzed using the pseudoaligner Salmon and differential expression quantified using DESeq2. Gene ontology analysis was then conducted using Panther. Pathway analysis was conducted using GeneAnalytics. Network analysis was conducted using the STRING plugin in the Cytoscape app. Results:- SCX knockdown had a larger effect on gene expression in fetal tenocytes, effecting 477 genes in comparison to the 183 genes effected in adult tenocytes, indicating that SCX-dependent processes may differ in these two developmental stages. Overall design: mRNA profiles of four independant lines of adult and fetal NT and shSCX expressing equine tenocytes were generated by RNA sequencing, using Illumina NovaSeq6000.