RNA-seq of cancer cell lines
A549 and UMUC3 cells were continuously cultured for a minimum of 30 days with CSC (25 microgram/ml) or an equivalent volume of DMSO vehicle control (0.0625%) before submitting for sequencing. Total RNA was extracted from cells treated with DMSO or CSC in three biological replicates per treatment. High RNA quality was verified for all samples (RNA integrity number equivalent, RINe>9.0) with Agilent ScreenTape. For each sample, 200 ng of total RNA was used to prepare an adapter-ligated library with the KAPA RNA HyperPrep kit with RiboErase using the xGen Dual Index UMI Adapters. The multiplexed libraries with 250-350 bp inserts were sequenced on a NovaSeq 6000 (Illumina) to generate 279-418 million paired-end 150 bp reads per sample.