(A) Heat map of 20 down-regulated genes and (B) 20 up-regulated genes in serine deprived bone marrow derived macrophages (BMDMs) stimulated with 100ng/mL LPS for 4 hours with or without extracelluar serine.
(C) IL-1β, n=6 (D) IL-10, n=6 (E) TNFα, n=3 mRNA expression in BMDMs stimulated with 100ng/mL LPS for 4 hours with or without extracellular serine. Data is normalized to LPS treated macrophages with serine and glycine.
(F) Protein secretion of IL-1β, n=6 (G) IL-18, n=7 (H) TNFα, n=4 in BMDMs primed with 100ng/mL LPS for 6 hrs followed by 5mM ATP for 30mins.
For (A-B), BMDMs (n=4) were stimulated with 100ng/mL LPS for 4 hours in the presence or absence of serine. See tables for complete RNAseq list of up-regulated and down-regulated genes in serine deprived BMDMs stimulated with LPS compared to control BMDMs stimulated with LPS.
For C-H, BMDMs were cultured in media with 400μM glycine, with or without 400μM serine, and supplemented with 1mM sodium formate where indicated. Data are shown as mean ± SEM. p values were calculated using a one-way paired ANOVA compared to LPS stimulated cells. *p<0.05.