GSM9407503: U2OS ZAK-KO + ZAKɑ Δ417-422 + Δ611-617 rescue, replicate 1; Homo sapiens; OTHER
Cells were subjected to 254 nm (UVC) irradiation with a dose of 150 mJ/cm². Cell pellets were lysed and sonicated using Branson Digital Sonifier. iCLIP2-seq libraries were prepared following the protocol described by Buchbender et al. (“Improved library preparation with the new iCLIP2 protocol,” Methods 178 (2020): 33–48), with adaptations inspired by Cordiner et al. (“Temporal-iCLIP captures co-transcriptional RNA-protein interactions,” Nat Commun 14(1):696, 2023). Cells were cultured in 15 cm dishes and harvested at ~80% confluence; two plates were used as starting material per condition. Final pooled libraries were size-selected for fragments ≥155 bp using AMPure XP beads (Beckman Coulter). Library quality was assessed with the Agilent High Sensitivity D1000 ScreenTape assay.