Expression profiling by high throughput sequencing (RNA sequencing)
Expression profiling by high throughput sequencing (RNA sequencing)
Cells were treated with accutase and counted. Approximately 1 million cells per sample were lysed. For all conditions, at least two biological replicate samples were collected from independent experiments.  Total RNA was extracted using the RNeasy Mini Kit according to the manufacturer's instructions (Qiagen). RNA concentration was quantified by NanoDrop, and quality was verified with a Fragment analyzer.  Library preparation was carried out using 0.5 μg of RNA with NEBNext Ultra II RNA Library Prep Kit (New England BioLabs, E7770S) as per the manufacturer's instructions. Libraries were amplified for 5 PCR cycles and purified with AMPure XP beads. DNA concentration was measured with a Qubit Fluorometer, and fragment size was determined with a Fragment analyzer.