GSM4727153: U2OS-GFP-Dcp1a_Mock2; Homo sapiens; RNA-Seq
Cells were treated with isotinic medium, hypertonic medium or resuced with isotonic medium after hypertonic treatment. Cells were harvested by scraping in RIPA buffer (Thermo-Fisher, PI89900). Total RNA was then extracted with QIAGEN RNeasy midi kit (Cat. No. 75144). RiboErase Beginning with at least 5 μg of total RNA, we first applied the Ribo-Zero rRNA Removal Kit (Illumina) to remove ribosomal RNA, and then proceeded with fragmentation, first- and second-strand synthesis, end repair, A-tailing, adaptor ligation, size selection, and uridine digestion. RNA integrity was measured on an Agilent 2100 Bioanalyzer using RNA Nano reagents (Agilent Technologies). Each sample was sequenced in duplicates using the Illumina HiSeq 2000 (with a 100-nt read length).