GSM5918247: RNA-IP, shADAR1 cells; Homo sapiens; RIP-Seq
Anti-HuR RNA-IP was carried out with a Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Cat. No. 17-700). Briefly, for each immunoprecipitation, 2 X 10^7 cells were harvested and lysed in 100 ul RIP Lysis Buffer with RNase and protease inhibitors. The lysate was subjected to immunoprecipitation using 5 ug anti-HuR antibody (Millipore, Cat. No. 03-102). The immunoprecipitated RNAs were extracted using phenol : chloroform : isoamyl alcohol (Thermo Fisher, Cat. No. AM9720) followed by quantitative PCR and RNA-seq. The RNA library was prepared using KAPA RNA HyperPrep kit (Roche, Cat. No. 07962312001).