Illumina HiSeq 4000 paired end sequencing; Total RNA-seq in the A549 cell line after glucocorticoid re-induction
Total RNA-seq in the A549 cell line after glucocorticoid re-induction
RNA was isolated from 1 million cells using the RNeasy kit from Qiagen. A549 cells were grown to approximately 90% confluence according to standard protocols in DMEM medium with 5% FBS. Cells were treated with either 100 nM dexamethasone or 0.1% ethanol. After 4 hours, cells were washed 2x with PBS and cultured in hormone-free medium for 24 hours, after which cells were treated again for 4 hours with either 100 nM dexamethasone or 0.1% ethanol. Ribosomal RNA was depleted using the KAPA RNA HyperPrep with RiboErase (Roche #08098131702). Sequencing libraries were prepared using KAPA RNA HyperPrep with RiboErase (Roche #08098131702).