ZFIN ID: ZDB-FIG-180620-30
Zhou et al., 2018 - MicroRNA-223 Suppresses the Canonical NF-κB Pathway in Basal Keratinocytes to Dampen Neutrophilic Inflammation. Cell Reports   22:1810-1823 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 4

NF-κB Pathway Is Elevated in Basal Epithelial Cells in miR-223−/− Embryos

The NF-κB reporter line Tg(NF-κB:GFP) was crossed into the miR-223−/− and matched WT background.

(A and B) Representative images (A) and quantification (B) of GFP signal. Mean fluorescence intensity (MFI) in the yellow square (A) in unwounded embryos or at the wound edge at 6 hpw was quantified. White arrows, neuromast cells constitutively expressing NF-κB signal.

(C) Quantification of GFP signal at the wound edge at 6 hpw in miR-223−/− embryos treated with DMSO or BAY.

(D) Representative confocal image of GFP+ cells in miR-223−/− embryos.

(E) Immunofluorescence of GFP and Tp63 (basal cell marker) in Tg(miR-223−/−, NF-κB:GFP) embryos. Nucleus were stained with DAPI. Representative confocal images of vertical view are shown in (E).

(F and G) Embryos from WT and miR-223−/− were injected with Pu.1 (200 μM) and Rac2 (100 μM) morpholinos. (F). Representative images showing the efficiency of the two morpholinos. Yellow arrowhead, macrophage; white arrowhead, neutrophils.

(G) Quantification of GFP signal at the wound edge at 6 hpw in WT and miR-223−/− embryos.

Scale bars, 100 μm (A and F), 20 μm (D and E). Data are representative of three independent experiments. ∗∗p < 0.01 and ∗∗∗∗p < 0.0001, unpaired Student’s t test (B and C) or two-way ANOVA (G). See also Figure S6 and Movie S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP mir223pu18/pu18; nc1Tg control Protruding-mouth caudal fin IFL
Protruding-mouth caudal fin epidermal basal stratum IHC
mir223pu18/pu18; nc1Tg transection: caudal fin Protruding-mouth caudal fin IFL
mir223pu18/pu18; nc1Tg + MO1-rac2 + MO1-spi1b transection: caudal fin Protruding-mouth caudal fin IFL
nc1Tg control Protruding-mouth caudal fin IFL
nc1Tg transection: caudal fin Protruding-mouth caudal fin IFL
nc1Tg transection: caudal fin, chemical treatment: NF-kappaB inhibitor Protruding-mouth caudal fin IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab7-tp63 IHC mir223pu18/pu18; nc1Tg control Protruding-mouth caudal fin epidermal basal stratum
Phenotype Details
Fish Conditions Stage Phenotype
mir223pu18/pu18; nc1Tg control Protruding-mouth caudal fin EGFP expression increased amount, abnormal
Protruding-mouth caudal fin I-kappaB kinase/NF-kappaB signaling increased process quality, abnormal
mir223pu18/pu18; nc1Tg transection: caudal fin Protruding-mouth caudal fin EGFP expression increased amount, abnormal
Protruding-mouth caudal fin I-kappaB kinase/NF-kappaB signaling increased process quality, abnormal
mir223pu18/pu18; nc1Tg + MO1-rac2 + MO1-spi1b transection: caudal fin Protruding-mouth caudal fin EGFP expression amount, ameliorated
Protruding-mouth caudal fin I-kappaB kinase/NF-kappaB signaling process quality, ameliorated
nc1Tg transection: caudal fin, chemical treatment: NF-kappaB inhibitor Protruding-mouth caudal fin EGFP expression decreased amount, abnormal
Acknowledgments:
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