ZFIN ID: ZDB-FIG-150612-4
Hall et al., 2013 - Immunoresponsive Gene 1 Augments Bactericidal Activity of Macrophage-Lineage Cells by Regulating β-Oxidation-Dependent Mitochondrial ROS Production. Cell Metabolism   18(2):265-278 Full text @ Cell Metab.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 4

JAK/STAT Pathway Cooperates with Glucocorticoid Receptor-Mediated Signaling to Regulate Infection-Responsive irg1 Expression within Macrophage-Lineage Cells

(A–C) Expression of irg1 within infected DMSO (control)-, AG490 (0.5 mM)-, and STAT3 inhibitor peptide (62.5 µM)-treated larvae, respectively, at 1 hpi.

(D) Quantification of irg1+ cells within individual infected larvae treated with DMSO, AG490 (0.5 and 1.0 mM), or STAT3 inhibitor peptide (62.5 and 125 µM), at 1 hpi (mean ± SD).

(E and F) Expression of irg1 within infected control MO-injected and Stat3-depleted larvae, respectively, at 1 hpi.

(G) Quantification of irg1+ cells within individual infected larvae as shown in (E) and (F) (mean ± SD).

(H and I) Expression of cebpβ within infected control MO-injected and Stat3-depleted larvae, respectively, at 1 hpi.

(J and K) Immunofluorescence detection of phosphorylated Stat3 (Stat3-P, Alexa 568) and Mpeg1/EGFP (Alexa 488) within the midbrain/hindbrain region of PBS-injected and infected Tg(mpeg1:EGFP) larvae, respectively, at 1 hpi. Yellow asterisks mark macrophage-lineage cells containing phosphorylated Stat3.

(L) Quantification of Stat3-P+ macrophage-lineage cells within the midbrain/hindbrain region of individual larvae as shown in (J) and (K) (mean ± SD). Region assessed, 100 µm from dorsal-most surface of midbrain/hindbrain (512 × 512, 50 z sections at 2 µm).

(M and N) Expression of irg1 within infected control MO-injected and Stat3 + Nr3c1-depleted larvae, respectively, at 1 hpi.

(O) Quantification of irg1+ cells within individual infected control MO-injected, Stat3-depleted, Nr3c1-depleted, and Stat3 + Nr3c1-depleted larvae, at 1 hpi (mean ± SD).

(P) Schematic illustrating proposed regulation of infection-responsive irg1 expression within macrophage-lineage cells. Asterisks, arrows, and arrowheads mark blood-specific, liver-specific, and gut-specific cebpβ expression, respectively. All views anterior to left. Numbers represent frequency of larvae with displayed phenotype. Scale bar, 100 µm in (A). p < 0.0001. See also Figure S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
acod1 WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment: tyrphostin B42 Long-pec macrophage ISH
WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec macrophage ISH
WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment: STAT3 inhibitor Long-pec macrophage ISH
WT + MO2-stat3 bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec macrophage ISH
WT + MO2-stat3 + MO4-nr3c1 bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec macrophage ISH
cebpb WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec blood macrophage ISH
Long-pec intestine ISH
Long-pec liver ISH
WT + MO2-stat3 bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec blood macrophage ISH
Long-pec intestine ISH
Long-pec liver ISH
EGFP gl22Tg control Long-pec macrophage IHC
gl22Tg bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec macrophage IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment: tyrphostin B42 Long-pec macrophage acod1 expression amount, ameliorated
WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec blood macrophage cebpb expression increased amount, abnormal
Long-pec macrophage acod1 expression increased amount, abnormal
WT bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment: STAT3 inhibitor Long-pec macrophage acod1 expression amount, ameliorated
WT + MO2-stat3 bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec blood macrophage cebpb expression increased amount, abnormal
Long-pec macrophage acod1 expression amount, ameliorated
WT + MO2-stat3 + MO4-nr3c1 bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium Long-pec macrophage acod1 expression amount, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Cell Metabolism for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

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.