RNA-seq of K562 xenograft: DFMO
Female NSG mice (Jackson: 005557) were injected subcutaneously with 3x106 K562 cells harboring the polyamine reporter construct. One day after injection, select mice were treated continuously with 1% DFMO (Eflornithine hydrochloride hydrate; MedChemExpress: HY-B0744B) in drinking water. Water bottles were exchanged every 5 days throughout the treatment window. Select mice were also treated daily with (3mg/kg) AMXT-1501 tetrahydrochloride (MedChemExpress: HY-124617A) formulated in sterile PBS and delivered intraperitoneally. All other experimental groups received daily injections of PBS as a vehicle control. Mice were treated for 14 days before doxycycline treatment to induce the polyamine sensor. Doxycycline hyclate (2mg/mL) was delivered in drinking water for 48 hours before tumors were harvested. RNA was isolated from tumors using TRIzol reagent. Libraries and sequencing were performed by Plasmidsaurus, Inc. Libraries for differential gene expression were prepared using BRB-seq, which employs barcoded poly-T primers to capture polyadenylated transcripts, which are reverse transcribed. The resulting double-stranded cDNA was tagmented and adapters were added during PCR amplification.
SUB16328046
DFMO_tumor_replicate_3