Supplementary Figure 7: Pharmacological inhibition of GPD2 modulates LPS activation and tolerance in BMDMs.
From: Glycerol phosphate shuttle enzyme GPD2 regulates macrophage inflammatory responses
(a) Seahorse extracellular flux analysis of maximal mitochondrial OCR in BMDMs stimulated with LPS for 1h +/- the GPD2 inhibitor iGP-1 (GPD2i), 300 μM (n=4). (b) ChIP-qPCR analysis of histone acetylation in Il6 and Il1b promoter regions in BMDMs treated as in a (n=3). (c) qPCR analysis of Il6 and Il1b gene expression in BMDMs stimulated with LPS for 0-3h +/- 300 μM GPD2i. (d) Seahorse analysis of maximal mitochondrial OCR in BMDMs stimulated with LPS for 12h +/- 300 μM GPD2i (n=3). (e) ChIP-qPCR analysis of histone acetylation in Il6 and Il1b promoter regions in unstimulated (U) or LPS-stimulated (U+LPS) naive BMDMs and unstimulated (T) or LPS-stimulated (T+LPS) tolerant BMDMs (n=3). Tolerance was induced by 24h LPS challenge +/- 300 μM GPD2i (T+LPS+GPD2i), which were both washed off before stimulation. (f) qPCR analysis of Il6 and Il1b gene expression in BMDMs treated as in e (n=3). Data are from one experiment representative of three independent experiments. Mean (a-f) +/- s.e.m. (a,d,f) shown. *p ≤0.05, **p ≤0.01, ***p ≤0.001 (two-tailed Student’s t-test).
