RNA was extracted from (RM1, 22Rv1, and LAPC4) under different conditions using RNeasy purification kit (74104, Qiagen) with DNAse treatment (79254, Qiagen). 50 ng was used for library preparation. The samples were then processed for RNA-sequencing using the NuGEN Universal RNA-Seq Library Preparation Kit with NuQuant (NuGEN Technologies, San Carlos, CA).  Briefly, 50 ng of RNA was used to generate cDNA and a strand-specific library following the manufacturer's protocol.