Figure 1.

Kinase Screen Reveals Distinct Metabolic Adaptation in Activated Macrophages

(A) Schematic of kinase screen.

(B and C) (B) Kinome modulation of LPS-induced (100 ng/ml; 1 hr) TNFα production (mean change ± SD) in RAW264.7 cells, including (C) kinases with known immune regulatory functions. Dotted lines indicate screen cut-off.

(D) 21 Kinases exceeded the screen cut-off. Blue bars represent protein kinases (PK) and white bars nonprotein kinases (NPK). Kinases involved in primary glucose metabolism are indicated by ^∗.

(E) Enrichment analysis for Canonical Pathways.

(F and G) (F) Bone marrow-derived or thioglycollate-elicited macrophages were stimulated with LPS (100 ng/ml) or (G) IL-4 (10 ng/ml), and ECAR and OCR were recorded.

(H and I) Dynamic (nonstationary) metabolic flux anaylsis of LPS or IL-4 stimulated BMDM by incubation with 100% labeled ¹³C-1-2-glucose. For abbreviations, see text. (H) shows isotope incorporation rate (m+n/total/10 min; m+n is all ¹³C-labeled molecules irrespective of mass shift, and total is all labeled and unlabeled), and (I) shows isotope distribution; ± SD, n = 5; ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.