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ZFIN ID: ZDB-FIG-160426-5
Berg et al., 2016 - Lysosomal Disorders Drive Susceptibility to Tuberculosis by Compromising Macrophage Migration. Cell   165:139-152 Full text @ Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 6
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagent:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 4

Cathepsin L Deficiency Causes snapc1b Mutant Vacuolated Macrophage Morphology and Susceptibility to M. marinum

(A) Quantitative real-time PCR of relative ctsl1 transcript in snapc1b+/- incross larvae at 6 dpf. Values normalized to transcript level of the heterozygous larvae, representative of two experiments.

(B and C) Confocal images of green fluorescent macrophages in larvae injected with red fluorescent MR-Cathepsin L at 3 dpf, either following treatment with E64d or DMSO control at 2dpf (B) or in snapc1b mutants and WT siblings (C). Yellow or white arrowheads denote macrophages that are positive or negative for MR-Cathepsin, respectively. Scale bar, 50 µm.

(D) Confocal images of green fluorescent macrophages stained with LysoTracker red in the brains of 3-dpf E64d-treated and DMSO control larvae. Scale bar, 50 µm.

(E) Average macrophage speeds during a 5-hr movie in the brains of 3-dpf E64d-treated and DMSO control larvae.

(F) Quantification of bacterial cording in DMSO control and E64d-treated larvae at 5 dpi with 150 Mm.

(G) Confocal images of green fluorescent macrophages stained with LysoTracker red in the brains of 3-dpf ctsl1 morphants and control larvae. Scale bar, 50 µm.

(H) Average macrophage speeds during a 5-hr movie in the brains of 3-dpf ctsl1 morphants and control larvae.

(I) Quantification of bacterial cording in control, snapc1b, and ctsl1 morphants at 5 dpi with 200 Mm.

(J) Quantification of vacuolated macrophages in the brains of 3-dpf WT or snapc1b mutant larvae following injection of ctsl1 RNA or control at 0 dpf.

(K) Quantification of bacterial cording at 2 dpi with 215 Mm in the HBV of snapc1b mutants and WT siblings following injection of ctsl1 RNA or control.

Statistical significance was assessed by ANOVA with Sidak’s post test (E, H, and J) or Fisher’s exact test (F, I, and K). See also Figure S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ctsl.1 snapc1bfh111/fh111(AB) standard conditions Day 6 whole organism RTPCR
YFP w200Tg + MO1-ctsl.1(AB) control Protruding-mouth macrophage IFL
w200Tg(AB) control Protruding-mouth macrophage IFL
w200Tg(AB) chemical treatment: aloxistatin Protruding-mouth macrophage IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
snapc1bfh111/fh111(AB) standard conditions Day 6 whole organism ctsl.1 expression decreased amount, abnormal
w200Tg + MO1-ctsl.1(AB) control Protruding-mouth macrophage lysosome accumulation macrophage, abnormal
Protruding-mouth macrophage morphology, abnormal
Protruding-mouth macrophage myeloid leukocyte migration decreased process quality, abnormal
Protruding-mouth macrophage myeloid leukocyte migration process quality, abnormal
w200Tg + MO1-ctsl.1(AB) bacterial treatment by injection: Mycobacterium marinum Days 7-13 response to bacterium decreased sensitivity of a process response to bacterium, abnormal
w200Tg(AB) chemical treatment: aloxistatin Protruding-mouth macrophage lysosome accumulation macrophage, abnormal
Protruding-mouth macrophage morphology, abnormal
Protruding-mouth macrophage myeloid leukocyte migration decreased process quality, abnormal
Protruding-mouth macrophage myeloid leukocyte migration process quality, abnormal
w200Tg(AB) bacterial treatment by injection: Mycobacterium marinum, chemical treatment: aloxistatin Days 7-13 response to bacterium decreased sensitivity of a process response to bacterium, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Cell, 165, Berg, R.D., Levitte, S., O'Sullivan, M.P., O'Leary, S.M., Cambier, C.J., Cameron, J., Takaki, K.K., Moens, C.B., Tobin, D.M., Keane, J., Ramakrishnan, L., Lysosomal Disorders Drive Susceptibility to Tuberculosis by Compromising Macrophage Migration, 139-152, Copyright (2016) with permission from Elsevier. Full text @ Cell