Female mice (8-12 week old C57Bl6) were injected i.t. with 1μg E.Coli derived (O127:B8) LPS in 50μl sterile saline, combined S.Aureus derived LTA (150μg) and PepG (50μg) or 1×106CFU of E. Coli (serotype ATCC25922) grown to log phase. At 24h post i.t. challenge, animals received either 30mg/kg AT7519 i.p. (10mg/kg for E.Coli experiments) in 200μl sterile saline or vehicle control. For wogonin experiments animals received 10mg/kg wogonin i.p. in DMSO or vehicle control. For experiments with bortezomib animals were pre-treated for 30min at 23.5h with 0.4mg/kg i.p. bortezomib in DMSO or vehicle control prior to administration of AT7519. At indicated time points lungs were lavaged with three aliquots of 800μl sterile ice cold saline. BAL fluid was centrifuged at 300g for 5min with supernatant from the first lavage stored for biochemical analysis. BAL total protein was measured by bicinchoninic acid assay (Pierce), BAL IgM by ELISA (eBioscience) and cytokines (TNF-α, IL-6, CCL-2, and IL-10) were measured by ELISA (R&D systems). BAL fluid cells were cytocentrifuged and stained using Diff-Quick™ and differential counts performed (≥450 cells counted per slide). Separate lungs for histology were fixed with 10% formalin (Sigma) prior to hematoxylin and eosin staining. For analysis of interstitial lung neutrophils, mice were perfused with 20ml PBS and the right lung digested in collagenase D (Roche) prior to incubation with anti-CD45 (Biolegend) and anti-Ly6G (Biolegend), with flow analysis in the presence of flow-check flurospheres (Beckman Coulter). Bacterial counts were determined following overnight incubation on LB agar plates while in vitro E.Coli growth in the presence of AT7519 was determined by absorbance (570nm) every 30min. Resolution intervals (Ri) were calculated as previously described 18 (link), 19 (link). Briefly, these were calculated as T50-Tmax, where Tmax is the time where neutrophil numbers were at their maximal numbers (Ψmax) and T50 is the time after Tmax where neutrophil numbers were half-maximal.