Fig. 3.

4-OI alleviates inflammation by inhibiting GAPDH activity. a, b RAW264.7 macrophages (a) and BMDMs (b) were treated with vehicle or 4-OI at the indicated concentrations. After 3 h, RAW264.7 macrophages or BMDMs were stimulated with LPS (1 μg/mL or 100 ng/mL) for 24 h, and IL-1β in the supernatants was measured by ELISA. c, d LPS-induced iNOS protein expression and its relative quantification after 24 h in RAW264.7 macrophages (c) and BMDMs (d) pretreated with or without 4-OI for 3 h. Data were corrected based on the actin loading control. e Treating LPS-stimulated RAW264.7 macrophages with 5 mM 2-DG, a glycolysis inhibitor, replicated the effect of 4-OI on IL-1β production. f IL-1β secretion was measured by ELISA in BMDMs that were treated with LPS ± DMF for 24 h in either limiting (0.5 mM) or saturating (25 mM) concentrations of glucose. Data shown are representative of three independent experiments. p Values were calculated using one-way ANOVA with Sidak’s correction or two-way ANOVA with Turkey’s correction for multiple comparisons tests. All data show mean ± SEM. Source data are provided as a Source Data file