GSM3703684: RIP MTR4 PLC/PRF5; Homo sapiens; RIP-Seq
RNA immunoprecipitation (RIP) was carried out with Magna RIP RNA-Binding Protein immunoprecipitation kit (Millipore; USA) according to the manufacturer's instruction. Briefly, cells overexpressing MTR4-HA were lysed by RIP lysis buffer containing protease inhibitor cocktail (Thermo fisher, USA) and RNase inhibitor (Invitrogen, USA). Nuclear pellets were collected and lysed by sonication. Nuclear lysates were incubated with anti-HA antibody or control IgG overnight at 4 °C, followed by incubation with protein A/G Dynabeads (Thermo fisher, USA) for 2 hours. After vigorous washing, pellets were resuspended in TRIzol (Invitrogen, USA). Co-immunoprecipitated RNAs were purified by RNAeasy Mini Kit (Invitrogen). RNA libraries were prepared for sequencing using standard Illumina protocols