GSM2837100: 3T3-L1; FLAG-tagged FTO (CLIP-Seq); Mus musculus; OTHER
After irradiation, cells were harvested and then lysed in RIPA lysis buffer (Sigma, R2078) at 4 ℃ for 2 h. Subsequently, the lysis mixture was centrifuged at 17,000 g at 4 °C for 30 min and the supernatant was carefully collected. 50 μl of antibody-conjugated Dynabeads Protein A was added into the supernatant and mixed gently at 4°C for 4 h. The supernatant was then discarded and the beads were washed for 3 times with 1 ml of high salt buffer and another 8 times with 1 ml of wash buffer (Wang et al., 2009). The RNAs were then isolated by proteinase K digestion following the manufacture's protocol. For CLIP-seq, NEB small RNA library kit
GEO Accession
GSM2837100