Subject	Molecular biology
Specific subject area	NGS, Transcriptomics
Type of data	Table
How data were acquired	RNA-Seq data acquired by Illumina NextSeq 500 (1 $\times$ 75 bp single-read sequencing)
Data format	Raw and processed data
Parameters for data collection	Total RNA was extracted and sequenced from human-telomerase-immortalized retinal pigment epithelial 1 (RPE-1) cells, treated with merbarone for 4 and 8 hours.
Description of data collection	Serum-starved human-telomerase-immortalized retinal pigment epithelial 1 (RPE-1) cells grown on 60mm plates were treated as required and total RNA was isolated with the RNeasy kit (QIAGEN, 74106), following instructions from the manufacturer. Then, total RNA (150ng) cDNA libraries were prepared using TruSeq Stranded mRNA (lllumina). Library size distribution was analyzed with Bioanalyzer DNA high-sensitive chip and Qubit. 1.4pM of each library was sequenced in NextSeq 500 HIGH-Output.
Data source location	Andalusian Molecular Biology and Regenerative Medicine Centre, Seville, Spain.
Data accessibility	RNA-Seq data (raw FASTQ, gene count tables and bigWig files) generated in this study are available under GEO accession number GSE198093. Lists of differentially expressed genes reported in this manuscript are available in the Supplementary Material.
Related research article	Andrés Herrero-Ruiz, Pedro Manuel Martínez-García, José Terrón-Bautista, Gonzalo Millán-Zambrano, Jenna Ariel Lieberman, Silvia Jimeno-González, Felipe Cortés-Ledesma Topoisomerase II$\alpha$ represses transcription by enforcing promoter-proximal pausing Journal DOI: doi.org/10.1016/j.celrep.2021.108977