ZFIN ID: ZDB-FIG-151117-4
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: Long-pec to Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Day 4

Fig. 4

ccr9a Is a Direct Target of Irf4a

(A) Co-expression of irf4a and ccr9a in the CHT of 2-dpf embryos revealed by double color FISH. Scale bar, 100 µm.

(B) qPCR of irf4a and ccr9a in GFP+ cells relative to negative cells sorted from coro1a:EGFP transgenic embryos at 2 dpf. p < 0.05; p < 0.01.

(C) Schematic diagram of the ChIP and reporter assay. The ccr9a-CF and CR are the control forward and reverse primers, which are used to amplify the ccr9a promoter region without the conserved Irf4 binding site. The ccr9a-1F and 1R represent primers used to amplify the promoter region with two conserved Irf4 binding sites within 500 bp upstream of 5′-ATG-3′. AICE, AP-1-IRF composite elements; EICE, Ets-IRF composite elements.

(D) Myc was detected in embryos injected with irf4a-myc mRNA by western blot.

(E) Direct binding of ccr9a promoter by Irf4a examined by ChIP assay.

(F) qPCR of DNA fragments obtained from ChIP assay with primers (ccr9a-1F, 1R or CF, CR). p < 0.001.

(G) Reporter assay showed the regulation of ccr9a promoter by Irf4a full-length CDS. M1 indicates that 5′-TGAT-3′ within AICE was mutated to 5′-GTCG-3′, and M2 indicates that 5′-GGAA-3′ within EICE was mutated to 5′-TTCC-3′. p < 0.01; p < 0.05.

(H) Expression of rag1 in the thymus (red arrow) of the control and ccr9a morphants at 4 dpf. All data are mean ± SD.

See also Figure S4 and Table S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
ccr9a WT control Long-pec intermediate cell mass of mesoderm ISH
irf4a WT control Long-pec intermediate cell mass of mesoderm ISH
rag1 WT control Day 4 thymus ISH
WT + MO1-ccr9a control Day 4 Not Detected thymus ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-ccr9a control Day 4 thymus rag1 expression absent, abnormal
Acknowledgments:
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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