ZFIN ID: ZDB-FIG-161202-13
Bernut et al., 2016 - Mycobacterium abscessus-Induced Granuloma Formation Is Strictly Dependent on TNF Signaling and Neutrophil Trafficking. PLoS pathogens   12:e1005986 Full text @ PLoS Pathog.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Day 5 to Days 7-13
PHENOTYPE:
Fish:
Condition:
Knockdown Reagents:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 5

IL8-dependent recruited neutrophils phagocytize Mabs at initial sites of infection.

(A) qRT-PCR measurement of il8 transcripts during Mabs R- or S-infections (≈150 CFU) in whole embryos at 2 and 5dpi. Mean log10 ± SEM of three independent experiments. (B) WT or il8 morphants Tg(mpx:eGFP) embryos were infected into the muscle (left), the hindbrain (middle) or the otic vesicle (right) with ≈100 R or S Mabs. Number of recruited neutrophils in injection sites at 3 hpi (two independent experiments, horizontal lines indicate the median values). (C) Microscopy showing representative neutrophil recruitment to R-abscesses in the brains of WT versus il8 morphants Tg(mpx:eGFP) larvae at 5 dpi. Scale bar, 100 μm. (D and E) WT or il8 morphants were iv infected with either R or S (tdTomato, ≈150 CFU). (D) Survival of R-(left) or S-infected (right) embryos (n = 50–60, average of two independent experiments). (E) Representative images of infected larvae at 3 dpi. Scale bars, 200 μm. (F and G) WT or csf3r morphants were iv infected with either R or S expressing tdTomato (≈150 CFU). (G) Survival of R-(left) or S-infected (right) embryos (n = 50–60, two independent experiments). (H) Representative images of infected larvae at 3 dpi. Scale bars, 200 μm. Statistical significance was determined by Kruskal-Wallis test with Dunns post-test (A), one-tailed unpaired Student’s t test (B) or by log-rank test (D and F).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cxcl8a WT bacterial treatment by injection: Mycobacteroides abscessus Day 5 whole organism RTPCR
Days 7-13 whole organism RTPCR
GFP i114Tg bacterial treatment by injection: Mycobacteroides abscessus Days 7-13 neutrophil IFL
i114Tg + MO4-cxcl8a bacterial treatment by injection: Mycobacteroides abscessus Days 7-13 neutrophil IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT bacterial treatment by injection: Mycobacteroides abscessus Protruding-mouth - Days 7-13 whole organism decreased life span, abnormal
Day 5 whole organism cxcl8a expression increased amount, abnormal
Days 7-13 whole organism cxcl8a expression increased amount, abnormal
WT + MO3-csf3r bacterial treatment by injection: Mycobacteroides abscessus Protruding-mouth - Days 7-13 whole organism decreased life span, abnormal
Day 5 response to bacterium decreased process quality, abnormal
WT + MO4-cxcl8a bacterial treatment by injection: Mycobacteroides abscessus Protruding-mouth - Days 7-13 whole organism decreased life span, abnormal
Day 5 response to bacterium decreased process quality, abnormal
i114Tg + MO4-cxcl8a bacterial treatment by injection: Mycobacteroides abscessus Protruding-mouth neutrophil chemotaxis decreased occurrence, abnormal
Protruding-mouth neutrophil immune response process quality, abnormal
Protruding-mouth neutrophil response to bacterium process quality, abnormal
Days 7-13 neutrophil chemotaxis decreased occurrence, abnormal
Days 7-13 neutrophil immune response process quality, abnormal
Days 7-13 neutrophil response to bacterium process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS pathogens for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Pathog.