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ZFIN ID: ZDB-FIG-170510-4
Kasper et al., 2017 - MicroRNAs Establish Uniform Traits during the Architecture of Vertebrate Embryos. Developmental Cell   40:552-565.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Prim-15 to Day 6
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-15 to Day 6

Fig. 5

miR-223 Does Not Regulate Phenotypic Variability of HSPC Number

(A) WISH of HSPC marker cmyb at transient sites of hematopoiesis, namely the DA ventral wall at 32 hpf, the caudal hematopoietic tissue between the DA and CV at 54 hpf, and the thymus, a definitive site of hematopoiesis, at 6 dpf (n = 10–20 embryos). Arrows show examples of cmyb+ cells. Yellow arrows indicate the region of the lateral trunk that is depicted in magnified images. Images captured with a 20× objective.

(B) Mean cmyb expression in miR-223Δ/Δ embryos normalized to actb1 levels and compared with wild-type as determined by qRT-PCR for three biological replicates. Bar plots show mean ± SEM, two-tailed Student's t test.

(C) Representative micrographs (25×) of the lateral trunk in embryos expressing Tg(kdrl:ras-mCherry)s896 and Tg(cmyb:GFP)zf169. Arrows point to examples of kdrl+cmyb+ cells budding from the DA ventral wall at the peak of hematopoiesis. Yellow arrows show region of the lateral trunk that is captured in zoomed-in images.

(D) Bar plots show the average of replicate means ± SEM of kdrl+cmyb+ cell number at 36 hpf (n = 4 replicates).

(E) Violin plots show kdrl+cmyb+ cell number probability density distributions. Solid lines in box plots depict median values. Phenotypic variability was not significantly different from wild-type, p = 0.24 by Levene's test (n = 36 embryos).

(F) Bar plots show average replicate SD ± SEM of kdrl+cmyb+ cell number (n = 4 replicates).

Significance calculations were relative to wild-type embryos. n.s., not significant (p > 0.05); p ≤ 0.05, ∗∗p ≤ 0.01, two-tailed Mann-Whitney U test unless otherwise indicated. Abbreviations as defined in Figure 1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP mir223ya304/ya304 ; s896Tg ; zf169Tg control Prim-25 hematopoietic multipotent progenitor cell IHC
Prim-25 ventral wall of dorsal aorta IHC
s896Tg ; zf169Tg ; mir223ya303/ya303 control Prim-25 hematopoietic multipotent progenitor cell IHC
Prim-25 ventral wall of dorsal aorta IHC
s896Tg; zf169Tg control Prim-25 hematopoietic multipotent progenitor cell IHC
Prim-25 ventral wall of dorsal aorta IHC
mCherry mir223ya304/ya304 ; s896Tg ; zf169Tg control Prim-25 dorsal aorta IHC
Prim-25 ventral wall of dorsal aorta IHC
s896Tg ; zf169Tg ; mir223ya303/ya303 control Prim-25 dorsal aorta IHC
Prim-25 ventral wall of dorsal aorta IHC
s896Tg; zf169Tg control Prim-25 dorsal aorta IHC
Prim-25 ventral wall of dorsal aorta IHC
myb AB control Prim-15 intermediate cell mass of mesoderm hematopoietic multipotent progenitor cell ISH
Long-pec hematopoietic multipotent progenitor cell ISH
Day 6 thymus hematopoietic multipotent progenitor cell ISH
mir223ya303/ya303 control Prim-15 intermediate cell mass of mesoderm hematopoietic multipotent progenitor cell ISH
Long-pec hematopoietic multipotent progenitor cell ISH
Day 6 thymus hematopoietic multipotent progenitor cell ISH
mir223ya304/ya304 control Prim-15 intermediate cell mass of mesoderm hematopoietic multipotent progenitor cell ISH
Long-pec hematopoietic multipotent progenitor cell ISH
Day 6 thymus hematopoietic multipotent progenitor cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
mir223ya303/ya303 control Prim-15 intermediate cell mass of mesoderm hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
Long-pec hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
Day 6 thymus hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
mir223ya304/ya304 control Prim-15 intermediate cell mass of mesoderm hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
Long-pec hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
Day 6 thymus hematopoietic multipotent progenitor cell myb expression increased amount, abnormal
mir223ya304/ya304 ; s896Tg ; zf169Tg control Prim-25 hematopoietic multipotent progenitor cell increased amount, abnormal
s896Tg ; zf169Tg ; mir223ya303/ya303 control Prim-25 hematopoietic multipotent progenitor cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 40, Kasper, D.M., Moro, A., Ristori, E., Narayanan, A., Hill-Teran, G., Fleming, E., Moreno-Mateos, M., Vejnar, C.E., Zhang, J., Lee, D., Gu, M., Gerstein, M., Giraldez, A., Nicoli, S., MicroRNAs Establish Uniform Traits during the Architecture of Vertebrate Embryos, 552-565.e5, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell