RNA-Seq of PC-3 cells grown at 18% O2
RNASeq was performed by Novogene. Briefly, mRNA was enriched using oligo (dT) beads for selecting polyA carrying RNAs and the Ribo-Zero kit was used for removing rRNA. RNA was subject to fragmentation before cDNA synthesis using random hexamer primers. Sequencing adaptor ligation, size selection, and PCR enrichment were performed to generate cDNA sequencing. The cDNA library was subject to QC processes, consisting of library concentration preliminarily tests with Qubit v2.0, insert size tests using Agilent 2100, and Q-PCR for library effective concentration precisely quantification. mRNA sequencing was performed on the Illumina NovaSeq 6000 S4 platform via PE sequencing at 150 bp x 2. Sequencing data was subject to QC for sequencing error rate, GC content, filtering by removing the adapters, and reads containing N >10% and low-quality bases with Q score < 5. The reads in the FASTQ file format from all samples were mapped to the Homo sapiens reference genome, GRCh38 from Ensemble (release 97) using STAR v2.7.0a