H1n1 pr8 34 strain

Primary mAECs, BMDMs or BMDCs were seeded into 48-well plates, grown to subconfluence, infected with H1N1 PR8/34 strain (Charles River) at 5 MOI, then incubated in the presence of DMSO, apoptosis or necroptosis pathway inhibitors for 24 or 48 h. Cell viability was assessed by MTS assay using CellTiter A_Queous one solution reagent according to the manufacturer's instructions (Promega, Madison, WI). Cell survival rate was calculated by comparison to DMSO-treated control cells and are presented as means ± SE (n = 3).

Age- and sex-matched littermate control and Runx3 general KO mice on C57BL/6 genetic background were anesthetized by i.p. injection of ketamine (100 mg/kg) and xylazine (8.5 mg/kg), and then given intranasally with 35 μL saline containing 1 LD₅₀ (~360 plaque-forming units (pfu)) H1N1 PR/8/34 strain or 35 μL saline alone as experimental controls. H1N1 PR/8/34 strain was from Charles River (Wilmington, MA, USA). These experiments were performed under a class II biosafety cabinet. Disposable PPE, such as gowns, gloves, masks, and shoe covers were used to prevent infection of personnel. Working areas exposed to live virus were disinfected with 70% ethanol, and hands were washed after experiments. The IAV-infected mice were housed in a designated BSL2 room and observed daily for signs of distress, body weight loss, and animal survival. Mice having a loss of more than 30% of their initial body weight were humanely euthanized and counted as dead. Bronchoalveloar lavage fluids (BALFs), lung, and mLN tissues were collected on days 3, 6, and 9 pi as we described [32 (link)].