design: RNAseq of HeLa and U2OS cells treated with either Nutlin, Darpin C10 targeting p53 or a control DARPin
library_construction_protocol: HeLa and U2OS cells were transfected with pcDNA3.1(+) plasmids containing the respective DARPin using Lipofectamin 2000 transfection reagent (Thermo Fisher Scientific) according to the manufacturer's instructions. Twenty-four hours after transfection cells were washed with PBS, detached and mRNA was isolated using the RNease mini kit (Qiagen). Cells were harvested and RNA was isolated using the RNeasy Mini Kit (QIAGEN, 74106) according to manufacturer's protocol. RNA was isolated using the RNeasy Mini kit (Qiagen) and RNA quality was assessed using the Experion RNA StdSens Analysis Kit (Bio‐Rad).  RNAseq libraries were prepared from total RNA with the Lexogen QuantSeq 3'-mRNA Library Prep Kit FWD for Illumina (Lexogen, 015.24) in combination with the UMI Second Strand Synthesis Module for QuantSeq FWD (Illumina, Read 1) (Lexogen, 081.96) following the manufacturer's protocol. Quality of sequencing libraries was controlled on a Bioanalyzer 2100 using the Agilent High Sensitivity DNA Kit (Agilent).