title: Illumina HiSeq 1500 sequencing; RNAseq of Calu-3 epithelial cells treated with Cyclosporin A
design: RNAseq of Calu-3 epithelial cells treated with Cyclosporin A
library_name: Mock1-24h_s
library_construction_protocol: Calu-3 cells were cultivated using DME/F12 with 10% FCS or DMEM with 10% DMEM.  Cells were infected with MERS-CoV at a multiplicity of infection (MOI) of 0.1 diluted in DMEM/F12 without FCS at 37°C for 1 h. Cells were washed with DMEM/F12 with 10% FCS and supplemented with stimulatory/ inhibitory reagents. Calu-3 cells were treated with either 10 µM Cyclosporin A (CsA, Sigma Aldrich) or DMSO solvent control. In addition  samples were treated with 20 µM calcineurin inhibitory peptide (CIP, Tocris), 10 µM SB203580 (Tocris), 10 µM SP600125 (Sigma Aldrich), 50 µM NFAT-inhibitor (Cayman Chemical Company).  Total RNA was isolated using RNA-Easy-Minikit followed by cDNA synthesis with random hexamer primers and RevertAid H Minus First Strand cDNA Synthesis Kit (ThermoFisher). RNA-seq libraries were prepared using the TruSeq Stranded mRNA Library Prep kit (Illumina).
attributes.tag: Experimental Factor: compound
attributes.value: dimethyl sulfoxide
attributes.tag: Experimental Factor: dose
attributes.value: 10