Anti ythdf3

Tumor cells seeded in a 10-cm dish at 80% confluence were cross-linked by ultraviolet light and harvested. RNA immunoprecipitation was performed with the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore) according to the manufacturer’s instructions. Cellular samples were subjected to RIP by anti-YTHDF3 (Santa Cruz) and mouse normal IgG (Millipore), which was served as negative control. Input and co-immunoprecipitated RNAs were extracted by TRIzol for further analysis by quantitative real-time PCR or high-throughput RNA-seq.

HBV 1.3-mer was a kind gift from Dr. Wang-Shick Ryu (Yonsei University) and obtained from the Addgene (Cambridge, MA, USA). FLAG-YTHDF1, 2, and 3 plasmids were a kind gift from Dr. Stacy M. Horner (Duke University Medical Center) (15 (link)). Antibodies were obtained as follow: anti-HBc antibodies from Dr. Haitao Guo (University of Pittsburgh, PA, USA) (19 ), anti preS2, anti-YTHDF3, anti-GAPDH antibodies from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-METTL3 antibody from Proteintech Group (Rosemont, IL, USA), anti-METTL14 antibody from Sigma-Aldrich, San Jose, CA, USA), anti-FTO and anti-YTHDF2 antibodies from Abcam (Cambridge, MA, USA), anti-FLAG, anti-PTEN, anti-IRF-3, and anti-phospho-IRF-3 antibodies from Cell Signaling Technology (Danvers, MA, USA).