Xpm1d3a25

For the Alternaria alternata extract (Alt Ext) model, mice were intranasally administered with 75 μL of 7.5 μg Alt ext (Greer Laboratories #XPM1D3A2.5) or PBS on days 0, 3, 6, and 9. Lungs were harvested on day 7 or 10 for endpoint analysis. For the house dust mite (HDM) model, mice were intranasally administered with 50 μL of either PBS or 100 μg of HDM Dermatophagoides pteronyssinus extract (Greer Laboratories #XPB70D3A25) plus 0.1 μg of LPS from Escherichia coli 0111:B4 (Sigma #L4391). Female mice were challenged on Day 0, 7, and 14 and cells and tissues were harvested on day 15 for all endpoints except for airway hyper-responsiveness measurements, which were performed on day 16. Inhibitor treatments were initiated on Day 0 and given daily throughout the 14 days of airway inflammation induction. Glut1 inhibitor (KL-11743, Kadmon Corporation) was dosed daily by oral gavage at 75 mg/kg as prescribed by the company dissolved in 0.5% methylcellulose and 0.25% Tween-80 (24 (link), 25 (link)). GLS inhibitor (CB839, Calithera Biosciences) was dosed twice daily via oral gavage at 200 mg/kg as described previously dissolved in 25% (w/v) hydroxypropyl- b-cyclodextrin in 10 mmol/L citrate, pH 2 (26 (link), 27 ). Dexamethasone was injected in the intraperitoneal daily for the last 5 days before harvest at 2.5 mg/kg.

Alternaria alternata extract (Greer Laboratories XPM1D3A25) was resuspended in PBS and concentration assessed by Pierce BCA assay (Thermo Fisher Scientific). HDM extract from Dermatophagoides pteronyssinus (Greer Laboratories XPB70D3A2.5) was resuspended in sterile PBS. Class IV Ovalbumin (Sigma, Gillingham, Dorset, UK) was lipopolysaccharide (LPS) depleted by Triton X-114 phase separation, and the levels of LPS present in HES and depleted OVA were below 0.1 and 0.01 IU LPS per mg protein, respectively by the Limulus Amoebocyte Lysate assay (Lonza, Slough, UK).