RNA-seq of SPARC-treated human corneal epithelial cells

We identified that Sparc gene expression is upregulated in corneal epithelial cells in a mouse model of dry eye disease involving lacrimal gland excision. Therefore, in this experiment we assess the effect of SPARC treatment on the transcriptome of human corneal epithelial cells. Overall design: We used a telomerase-immortalized human corneal epithelial cell line (hTCEpi) that was graciously donated by Danielle Robertson (UT Southwestern). These cells were maintained in keratinocyte growth medium 2 (PromoCell C20011) supplemented with SupplementMix, 1% penicillin/streptomycin, and 0.15 mM calcium chloride. We treated hTCEpi cells with recombinant human SPARC (PeproTech 120-36) in KGM media: 0.00, 0.05, 0.50, and 5.00 micrograms/ml for 24h. Then, we washed cells with PBS, isolated RNA, and prepared samples for RNA-sequencing using the RiboErase rRNA depletion kit.