Np gtx125989

The antibodies used for immunoblotting include: mouse monoclonal antibodies (mAbs) c-Myc antibody (sc-56634) and β-actin (C4) (sc-47778) (both purchased from Santa Cruz Biotechnology, Dallas, TX, USA); rabbit polyclonal antibodies against NP (GTX125989) was purchased from GeneTex (Taiwan); anti-TBK1 (ab40676), and anti-TBK1 (phosphor S172) (ab109272) antibodies were obtained from Abcam (Cambridge, MA, USA). Rabbit polyclonal antibody against acetylated-lysine (9441), mouse mAb against HDAC1 (10E2) (5356), p-IRF3 (4947), IRF3 (4302), p-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (4370), p44/42 MAPK (Erk1/2) (9102), and rabbit mAb HA-Tag (c29F4) (28) were from Cell Signaling Technology (Beverly, MA, USA).

Hit compounds were purchased from Namiki Syoji Co. Ltd (Tokyo, Japan) and dissolved in 100% dimethyl sulfoxide (DMSO). Benzbromarone was purchased from Tokyo Chemical Industry Co. Ltd (Tokyo, Japan) and dissolved in 100% DMSO. Oseltamivir phosphate purchased from F. Hoffmann-La Roche Ltd (Basel, Switzerland) was dissolved in phosphate-buffered saline (PBS) at a concentration of 10 mM. All of the compounds were maintained at −30 °C until use. Before performing the experiments, the compounds were diluted with Eagle’s minimum essential medium (MEM) supplemented with 1% 100× vitamin solution (MEM vitamin). HeLa cells were obtained from Dr Takujiro Homma (Nagasaki University, Japan) and maintained in Dulbecco’s modified Eagle’s medium (Sigma Aldrich, St Louis, MO) containing 10% fetal bovine serum (FBS). MDCK cells kindly donated by Dr Kyosuke Nagata (Tsukuba University, Japan) were grown in MEM supplemented with 5% FBS. These cells were maintained at 37 °C in an atmosphere of 5% CO₂. Influenza viruses were prepared as described previously^{48 (link)} and stored at −80 °C. Anti-HA (GTX127357), -M1 (GTX125928), -PA (GTX118991), -PB1 (GTX125923) and -NP (GTX125989) antibodies were purchased from GeneTex, Inc. (Irvine, CA). Anti-actin (A5060) antibody was purchased from Sigma Aldrich.