title: GSM8239473: HEK293T cells, DHX29-IP, rep2; Homo sapiens; OTHER
library_name: GSM8239473
library_construction_protocol: Cells were transfected with the pFLAG-DHX29-WT vector, and the transfected cells were subjected to in vivo DSP crosslinking. After that, the lysates were treated with RNase I, followed by the treatment of SUPERase•In RNase inhibitor. Some of the lysates were taken out and used for input. The lysates were then layered on the sucrose cushion and subjected to ultracentrifugation. The supernatants were discarded, and the pellets were resuspended in the Lysis buffer. The samples were pre-cleared with Dynabeads Protein G, followed by the incubation with Dynabeads Protein G pre-incubated with the anti-FLAG antibody. Then, the beads were washed and mixed with TRIzol (ThermoFisher Scientific). Thor-Ribo-Seq was performed as described previously (Mito et al., bioRxiv. 2023).