Ab167264

Cell lysates were resolved using 12% SDS‐PAGE. Anti‐EHF (Abcom, ab167264), anti‐p‐STAT3 (SAB4300033‐100UG, rabbit polyclonal), anti‐STAT3 (SAB2102317‐100UL, rabbit polyclonal), and anti‐ACTB (ACTN05 [C4]) antibodies were used as primary antibodies. Protein‐blotted membranes were incubated with antibodies using Can Get Signal Immunoreaction Enhancer Solution (Toyobo) at 4℃ overnight for the primary antibodies and at room temperature for 1 hour for secondary antibodies, followed by visualization using the ECL Prime system (GE Healthcare). The protein signals were detected using LAS‐3000 (Fujifilm).

Clinical GC tissue samples were obtained from patients undergoing gastrectomy at Tokyo University Hospital, with written informed consents. Formalin‐fixed, paraffin‐embedded tissue blocks of EBV(−) and EBV(+) GC were cut into 4‐μm‐thick sections. Three serial sections were prepared for each tissue block and used for hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) using anti‐EHF (Abcom, ab167264) and anti‐p‐STAT3 (SAB4300033‐100UG, rabbit polyclonal) antibodies. IHC was performed using Ventana BenchMark automated immunostainer (Ventana Medical Systems), and visualized with OptiView DAB IHC Detection Kit (760‐700; Roche) according to the manufacturer's instructions. Two independent experienced pathologists scored the staining as follows: 3 (strong), 2 (moderate), 1 (weak), and 0 (negative). The study design was approved by the Institutional Review Board of Chiba University and the University of Tokyo.