RIP-seq for HNRNPD overexpression in nasopharyngeal carcinoma.

RNA immunoprecipitation assays were performed using the RNA Immunoprecipitation Kit (Bersinbio, Bes5101, China) according to the manufacturer provided protocol. Briefly, HK1-R cells transfected with 3HA-HNRNPD were irradiated and lysed in RIP buffer containing protease and RNase inhibitors to prevent DNA contamination. The cell lysis samples were divided into thirds according to 0.8 mL (IP), 0.8 mL (IgG), and 0.1 mL (Input). The IP and IgG samples were incubated overnight at 4 degrees C with 1 ug each of anti-HA antibody and IgG antibody, respectively. Equilibrated protein A/G beads were added and incubated at 4 degrees C for 1 h. RNA was then extracted by the Trizol-chloroform method and analyzed by RIP-seq.