Analysis of the senescence transcriptome after knockdown of AHCY

IMR90 ER:RAS cells were infected with 2 different lentiviral pGIPZ shRNAs against AHCY or with an empty vector (EV). We treated IMR90 ER:RAS cells with 4OHT to induce RAS activation or with vehicle (DMSO) as a non-senescent control. RNA was collected 6 days after the start of the experiment once the senescence program has been fully implemented. Our previous experiments showed that knockdown of AHCY prevents senescence. By performing RNAseq, we have unveiled that AHCY knockdown prevents upregulation of a set of genes essential for the onset of senescence and its associated secretory phenotype. 16 samples examined from 4 different treatment (4 replicates each): non- senescent cells (EV), senescent cells (EV+4OHT), shAHCY_2 (+4OHT), shAHCY_4 (+4OHT).