GSM5643611: G7_6hrs_THZ531_repl1; Homo sapiens; RNA-Seq
SLAM-seq was performed according to the protocol by reference. Briefly, cells were seeded at approximately 70% confluency, and treated with THZ531 or vehicle control. 1 hour before harvest, 4sU was added to the medium at a final concentration of 500 μM. The samples were kept in the dark as RNA was extracted with Qiagen's RNeasy Plus Mini Kit. 3 μg RNA was alkylated with iodoacetamide (Sigma, 10 mM) for 15 minutes, and the RNA was repurified by ethanol precipitation. 250 ng RNA was used to make libraries with Lexogen's QuantSeq 3′ mRNA-Seq Library Prep Kit FWD for Illumina and PCR Add-on Kit for Illumina. Deep sequencing was performed using the NovaSeq platform (Illumina).