Genomewide transcriptional analysis of growth hormone-treated human podocytes

The growth hormone plays a significant role in normal renal function and overactive growth hormone signaling has been implicated in proteinuria in diabetes. Earlier studies from our group have shown that the glomerular podocytes, which play an essential role in renal filtration, express the growth hormone receptor, suggesting the direct action of growth hormone on these cells. Nevertheless, the precise mechanism and the downstream pathways that are induced by the excess growth hormone in these podocytes leading to diabetic nephropathy are not clearly established. To compressively understand the growth hormone’s effect on podocytes at transcript level we performed RNA-Sequencing. Conditionally immortalized human podocytes were employed in this study. Human podocytes in duplicates were treated with (500 ng/ml) or without growth hormone for 30 min and total RNA was isolated. Both by running on an agarose gel and using the bioanalyzer confirmed the quality of RNA. High-quality RNA libraries were constructed using mRNA templates, random hexamers, oligo-dT primers, and Illumina second strand synthesis buffer. RNA sequencing was performed on Illumina 2500 Hi-Seq devise by pair-end reads at Nucleome Informatics Facility, Hyderabad, India.