Effect of Transforming Growth Factor-beta 1 on the Rat Renal Mesangial Cell Transcriptome

Rat renal mesangial cells (SciencCell, #R4200) were grown to sub-confluence on poly-L-lysine coated flasks in mesgnaial cell medium (SciencCell, #R4201) supplemented with 2�S, growth supplement (SciencCell, #R4252) and pen/strep. Cells were quiesced for 48h and then treated for 24h with human recombinant TGF-beta 1 (5 ng/mL; Sigma#T7039) in complete media (n=4) or fresh media without growth factor (n=4). Total RNA was extraced from cell pellets using standard protocols. Whole RNA stranded libraries were prepared using Illumina's Ribo-zero Gold protocol and sequenced on an Illumina NovaSeq platform (150bp PE). After demultiplexing and quality control, the data was then processed through RNA-seq expression analysis workflow, which included trimming, alignment, transcript assembly, feature quantification and, differential expression analysis with edgeR (verison 3.38.4).