ZFIN ID: ZDB-FIG-151117-1
Wang et al., 2015 - Irf4 Regulates the Choice between T Lymphoid-Primed Progenitor and Myeloid Lineage Fates during Embryogenesis. Developmental Cell   34(6):621-31 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 1

Irf4 Is Expressed in Lymphoid Progenitors at Embryonic Stages

(A) Expression pattern of irf4a examined by WISH. The expression of irf4a was observed in the CHT at 2 dpf and later in the thymus at 3 dpf. Red arrows indicate irf4a-expressing cells. The magnified view of the boxed region is shown in the right panel, and the thymus is denoted with red circle.

(B) Absence of irf4a expression in the CHT of the cloche mutants and runx1 morphants.

(C) qPCR of irf4a in sorted cells from 3-dpf transgenic embryos. Irf4a is highly expressed in cmyb:EGFP+ cells relative to negative cells but is not expressed in mpo:EGFP cells or gata1:GFP cells. The purity of mpo:EGFP cells or gata1:GFP cells was validated by the high expression level of lyz or gata1, respectively, with β-actin as the control. p < 0.001. nd, not detectable.

(D) Expression of irf4a in a subset of cmyb-expressing cells in the CHT revealed by double color FISH. The cell denoted by the white arrow is shown in the insets with a magnified view. Scale bar, 20 µm.

(E) Western blot showed the protein level of IRF4 in the FL (E11.5, E12.5, and E14.5) or thymus (E16.5) of mouse embryos.

(F) Expression of IRF4 in the FL of E14.5 mouse embryos examined by immunochemistry. Red arrows indicate IRF4-expressing cells. Scale bar, 200 µm.

(G) qPCR analysis of the relative mRNA level of Irf4 in purified hematopoietic cells from the FL of E13.5 mouse embryos.

(H) Expression of IRF4 was observed in CCR9+ cells sorted and photographed using FlowSight (Merck-Millipore). Scale bar, 20 µm. All data are mean ± SD.

See also Figure S1 and Table S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
gata1a la781Tg standard conditions Protruding-mouth erythroid lineage cell RTPCR
irf4a WT control Prim-5 Not Detected trunk ISH
Long-pec hematopoietic multipotent progenitor cell ISH
Long-pec intermediate cell mass of mesoderm ISH
Protruding-mouth hematopoietic system ISH
Protruding-mouth thymus ISH
WT standard conditions Protruding-mouth hematopoietic system ISH
WT + MO1-runx1 standard conditions Long-pec Not Detected intermediate cell mass of mesoderm ISH
i114Tg standard conditions Protruding-mouth Not Detected granulocyte RTPCR
la781Tg standard conditions Protruding-mouth Not Detected erythroid lineage cell RTPCR
npas4lm378/m378 standard conditions Protruding-mouth Not Detected hematopoietic system ISH
zf169Tg standard conditions Protruding-mouth hematopoietic multipotent progenitor cell RTPCR
lyz i114Tg standard conditions Protruding-mouth granulocyte RTPCR
myb WT control Long-pec hematopoietic multipotent progenitor cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-runx1 standard conditions Long-pec intermediate cell mass of mesoderm irf4a expression absent, abnormal
npas4lm378/m378 standard conditions Protruding-mouth hematopoietic system irf4a expression absent, 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, 34(6), Wang, S., He, Q., Ma, D., Xue, Y., Liu, F., Irf4 Regulates the Choice between T Lymphoid-Primed Progenitor and Myeloid Lineage Fates during Embryogenesis, 621-31, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell