Identification of target RNAs regulated by HuR

In this work, we determined RNA degradation rate of whole transcriptome in HeLa transfected with siRNA against HuR, an AU-rich RNA binding protein. The knockdown cells were cultured under with 150 micro molar BrU for 24h. Then medium was replaced with normal growth medium, DMEM containing 10% FBS and antibiotics, to wash out BrU. After changing medium, cells were collected and total RNAs labeled with BrU were isolated by using RNAiso Plus reagent. BrU labeled RNAs were isolated by specific mouse monoclonal antibody against BrU, clone 2B1. Finally, isolated BrU labeled RNAs were analyzed by illumina platform.