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Fig. 7

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ZDB-FIG-150612-7
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Hall et al., 2013 - Immunoresponsive Gene 1 Augments Bactericidal Activity of Macrophage-Lineage Cells by Regulating β-Oxidation-Dependent Mitochondrial ROS Production
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Fig. 7

Irg1 Is Required for Enhanced mROS Production within Macrophage-Lineage Cells in Response to Fatty Acid Supplementation

Inhibition of β-oxidation reduces LPS-stimulated mROS production and bactericidal activity of J774.2 murine macrophage-like cells.

(A and B) Live confocal imaging of mROS production by macrophage-lineage cells within the midbrain/hindbrain region of control MO-injected Tg(mpeg1:EGFP) larvae, at 3 hpi, following Salmonella (Sal.)/MitoSOX injection alone or supplemented with an equimolar fatty acid mixture, respectively. MitoSOX fluorescence intensity is displayed as a colormap, with warmer colors representing higher signal intensities.

(C) Quantification of mROS production by macrophage-lineage cells (measured as total fluorescence intensities of MitoSOX within individual mpeg1+ cells), as marked in (A) and (B), within control MO-injected and Irg1-depleted Tg(mpeg1:EGFP) larvae at 3 hpi, following the indicated treatments (mean ± SD). Five to ten larvae were sampled for each treatment.

(D) Quantification of MitoSOX fluorescence, as detected by flow cytometry (measured as mean fluorescence intensity), within J774.2 murine macrophage-like cells treated with LPS alone (500 ng/ml) or supplemented with 50 µM etomoxir, compared to untreated, n = 4 biological replicates (mean ± SD).

(E) Gentamicin protection assay showing infected DAPI-stained J774.2 cells (either untreated (control) or treated with 50 µM etomoxir) at 2, 8, and 24 hr postinfection (hpi) with Sal-GFP.

(F) Quantification of recovered Sal-GFP following cell lysis and plating of samples, as shown in (E), n = 3 biological replicates (mean ± SD).

(G) Model for Irg1/β-oxidation-dependent bactericidal mROS production. Scale bars, 5 µm in (A) and (E). Abbreviations: n.s., not significant; p < 0.05; p < 0.001; p < 0.0001. See also Figure S7.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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Reprinted from Cell Metabolism, 18(2), Hall, C.J., Boyle, R.H., Astin, J.W., Flores, M.V., Oehlers, S.H., Sanderson, L.E., Ellett, F., Lieschke, G.J., Crosier, K.E., and Crosier, P.S., Immunoresponsive Gene 1 Augments Bactericidal Activity of Macrophage-Lineage Cells by Regulating β-Oxidation-Dependent Mitochondrial ROS Production, 265-278, Copyright (2013) with permission from Elsevier. Full text @ Cell Metab.