*2.2. Bioinformatics Studies*

#### 2.2.1. Quality Assessment of RNA-seq Data, Mapping and Read Annotation

All processing and analysis on raw data were performed using Ubuntu 20.00 (64-bit) and open-source software available through the R/Bioconductor. After check and quality control of paired-end reads with the final version of MultiQC (https://github.com/ewels/ MultiQC, 25 January 2022) and Trimmomatic (http://www.usadellab.org/cms/?page= trimmomatic, 25 January 2022), the remaining reads as clean reads were mapped to the genome reference GRCh37 (hg19) using the star (https://github.com/alexdobin/STAR/ releases, 10 January 2022) package, and mapping efficiencies accounted for 98.50%. The counting of transcripts was also performed with Htseq-count (https://htseq.readthedocs. io/en/release\_0.11.1/count.html, 10 January 2022).
