Anti gpd2

For immunoblotting, BMDMs were lysed in 1% NP-40 buffer or RIPA buffer, and protein concentration was determined using the Bradford method or micro BCA assay (Pierce). Primary antibodies used were anti-phospho-ACLY(S455) and anti-ACLY (1:1000, Cell Signaling), anti-GPD2 (1:800, ProteinTech), anti-IκB (1:1000, Santa Cruz), anti-phospho-IRF3 (1:1000, Cell Signaling), and anti-α-tubulin (1:5000, Sigma-Aldrich). For ELISA, cell culture supernatants and mouse whole blood were spun down (5 min at 400 g and 20 min at 13,000 g, respectively), diluted appropriately in 5% BSA, and assayed for IL-6 using BioLegend Standard ELISA kits according to the manufacturer’s protocols. For in vitro LPS tolerance experiments, cell culture supernatants in the T and T+L conditions were collected after LPS washout and rest or restimulation for 4h (refer to Fig 5a; time of collection indicated by “Assay”), allowing for measurement of IL-6 produced during tolerance rather than activation. Supernatants were analyzed from duplicate or quadruplicate cell cultures, indicated by n in figure legends. Full scans of western blots can be found in Supplementary Fig. S9–11.