|ZFIN ID: ZDB-FIG-151117-4|
|Stage Range:||Long-pec to Day 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.
|ccr9a||WT||control||Long-pec||intermediate cell mass of mesoderm||ISH|
|irf4a||WT||control||Long-pec||intermediate cell mass of mesoderm||ISH|
|WT + MO1-ccr9a||control||Day 4||Not Detected||thymus||ISH|
|WT + MO1-ccr9a||control||Day 4||thymus rag1 expression absent, abnormal|
This image is the copyrighted work of the attributed author or publisher, and
ZFIN has permission only to display this image to its users.
Additional permissions should be obtained from the applicable author or publisher of the image.
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