Fig 2: NLRP3 activation alters macrophage glycolysis causing a decrease of ATP in S. aureus(A) Glucose uptake into BMDMs was measured at 24 hpi using the Glucose Uptake-Glo assay kit (Promega). BMDMs were primed for 4h with 10 ng/ml LPS, followed by treatment with 10 μM MCC950 or vehicle (DMSO) for 45 min and infection at MOI 10 with WT LAC. Data were normalized to RLU from uninfected samples.(B) Seahorse XF Glycolysis Stress Assay was used to measure glycolysis from BMDMs untreated or treated with 100 μM MCC950 followed by stimulation with 50 ng/ml alpha toxin.(C) ATP levels in S. aureus as measured by luminescence. BMDMs treated with and without MCC950 prior to infection at MOI 10 with LACluxABDCE. Luminescence was measured and normalized to CFU. See also Figure S4. Statistical significance was determined using one-way ANOVA with Sidak’s multiple comparison (A, B) or using an unpaired t-test (two-tailed) (C). All experiments were performed in biological triplicate at least twice on two separate days (n ≧ 6). Bars represent the mean +standard deviation. See also Figure S4.