Total RNA-Seq in KSHV lytic reactivation with spliceosome inhibition

Subconfluent monolayers of iSLK-BAC16 were induced with 1 ug/mL Doxycycline, 1 mM Sodium Butyrate in 1x DMEM media supplemented with 2% Tet-approved FBS. 0 hour time point was when induction media was added and cells were first placed at 37 degrees Celsius to incubate. If indicated, cells were treated with major spliceosome inhibitors at 4 hours pre- or 24 hour post-induction. At these times, media was removed from infected cells and replaced with induction media containing 30 uM Isoginkgetin (CAS #548-19-6, Sigma #416154), 25 nM Pladienolide B (CAS #445493-23-2, SantaCruz #sc-391691), or vehicle (DMSO, 0.1%). Total RNA was collected at 24 or 48 hours post induction using the Direct-zol RNA MiniPrep Kit. ERCC spike-in controls were added to total RNA. RNA was ribominus selected and directional cDNA libraries were generated. 2 biological replicates were sequenced for all samples. Sequencing was performed using the Illumina HiSeq 4000 or Illumina NovaSeq SP platform to generate 150 bp PE reads.