SRSF2 transcriptional function maintains genome integrity during cell division [cell lines]

To investigate the molecular mechanisms through which SRSF2 acts it was depleted in 3 epithelial cell lines; primary human keratinocytes (HK) and two squamous cell carcinoma cell lines SCC25 and FaDu cells. Additionally FaDu cells were used to make clones with stably reduced SRSF2 levels in order to explore how cells can compensate reduced SRSF2 levels. Due to SRSF2's role in transcription Cut&Run using an antibody specific for total Pol II was performed in ctr and SRSF2 depleted FaDu cells. Overall design: SRSF2 depletion was achieved using an siRNA pool. Total RNA was isolated 48 hours after transfection, using Trizol reagent and RNA-sequencing libraries were prepared. SLAM-sequencing and Cut&Run were also performed 48 hours after transfection and libraries were prepared from Total RNA that had undergone thiol modification or on DNA that was pulled down. RNA was also isolated from stable clones and a control clone and RNA-sequencing libraries were prepared.