GSM8326618: hSAECs, OGG1, RNA-IP, Rep-1; Homo sapiens; RIP-Seq
At 24 hpi, cells were fixed with 1% formaldehyde for 10 min at room temperatrue. After quenching and cell lysis by RIPA, lysates were briefly sonicated. During the preparation process, we employed the iron chelator deferoxamine mesylate (252750, Millipore-Sigma) to prevent additional oxidation. The supernatant was treated with RNase A (20 µg/mL) and RNase T1 (5U) at 37°C for 30 min to digest naked viral RNA. We then subjected the supernatant to IP with an anti-OGG1 antibody (PA5-86046, Invitrogen) for 4 hours, using head-over-tail rotation for effective mixing. We incubated the ribonucleoprotein-antibody complexes with magnetic beads for 1 hour at 4°C and subsequently washed them five times with RIPA buffer. After proteinase K digestion and reversing the crosslinks, we extracted the RNA samples using TRIzol. The libraries were processed for Illumina sequencing. stranded RNA-seq library