mRNA expression profiling of DNA damage-induced senescent cells

To identify differentially expressed genes during cellular senescence, we performed transcriptional profiling with RNA sequencing (RNA-seq) before and after senescence induction using a stress-induced senescence model (bleomycin-induced senescence). IMR90 cells were treated with bleomycin (10 ug/ml for 24 h) to induce senescence, and total RNAs from normal (-bleomycin) and senescent (+bleomycin, 10 days after exposure to bleomycin) IMR90 cells were isolated using the acidic phenol extraction method. One microgram of high-quality RNA samples (RIN > 7.0) was used to construct RNA-seq libraries using the TruSeq Stranded mRNA LT Sample Prep Kit (Illumina). RNA-seq was performed on an Illumina NovaSeq 6000 sequencer.