title: NextSeq 500 sequencing; Deficiency of ERO1 and activation of protein translation synergistically impair breast tumor resilience
design: Deficiency of ERO1 and activation of protein translation synergistically impair breast tumor resilience
library_construction_protocol: Genetic modification, ERO1-LÎ± CRISPR-Cas9 Knock-out MDAMB231 metastatic breast tumors. Collection of 5-10 million cells in PDS, and resuspension of the pellet in a buffer. Paclitaxel was injected intravenously (i.v.) at the dose of 15 mg/kg, Q7x2 (every seven days for 2 weeks) and stopped two weeks before the end of the experiment. ISRIB (trans-isomer, Aurogene s.r.l) was dissolved in 10% DMSO (Sigma-Aldrich) in corn oil. It was injected interaperitoneally (i.p.), at the dose 2.5 mg/kg every two days for three weeks. Forty-eight hours after the last dose of ISRIB, mice were sacrificed, and primary breast tumors were isolated and RNA extracted. Protocol: miRNeasy Mini Kit TruSeq Stranded Total RNA Sample Preparation Guide