4 eBiosciences, San Diego, CA), or anti-TCRβ (clone H57-597, BD Pharmigen) Allophycocyanin, PE-, or FITC-conjugated mAb and analyzed with a FACS-Calibur (BD Pharmingen). Data were analyzed using the FlowJo program (Tree Star, Ashland, OR). We thank Dr. Nancy Andrews for providing us with the mHfe-C282Y KI mice, Dr. Hélène Coppin and Marie-Paule Roth for providing us with the DBA/2 mHfe KO mice, Dr. Benedita Rocha for providing us with H-2 Db-restricted anti-HY TCR transgenic Rag 2 KO male mice, Prof. Albert Bendelac for providing alpha-GalCer CD1d tetramer, Prof.
Christian Boitard for constructive support of the project, Prof. Terry Delovitch for careful reading of the manuscript, Christine Detchepare for excellent secretarial assistance Navitoclax mouse and Frances Sheppard for careful proofreading. This work was funded by the Institut Pasteur, the Institut National de la Santé et de la Recherche Médicale and the Ligue contre le Cancer du Grand Est. Rachid Boucherma was supported by the Fondation Transplantation. Hedia Kridane-Miledi was supported by the Ministère de l’Enseignement Supérieur et de la Recherche. François A. Lemonnier, Selleck Idelalisib Pierre-Simon Rohrlich, and François Huetz shared seniorship. Conflict of interest: The authors declare
no financial or commercial conflict of interest. Disclaimer: Supplementary materials have been peer-reviewed but not copyedited. Supporting information Figure 1 Gating Strategy and CD4/CD8 plot of TCRβ positive cells Supporting information Figure 2 PLZF expression in mHfe WT, Rag 2 KO α+/−β+/− anti-mHFE TCR-transgenic DBA/2 mouse T lymphocytes and purified DBA/2 WT NKT cells Supporting information Figure 3 Cytokine and hepcidin productions by H-2 Db-restricted anti-HY TCR transgenic T lymphocytes from male mice “
“Cellular differentiation of the T-cell branch of the immune system begins with the HSC, which undergoes a series of stages characterized by progressive restriction
in multipotency and acquisition of specific lineage identity At the molecular level, the restriction of cell potential, commitment, and differentiation to a specific lineage is achieved through the coordinated control of gene expression check and epigenetic mechanisms. Here, we analyzed and compared the gene expression profiles and the genome-wide histone modification marks H3K4me3 (H3 lysine 4 trimethylation) and H3K27me3 (H3 lysine 27 trimethylation) in (i) in vitro propagated HSCs, (ii) in vitro generated and propagated pro-T cells derived from these stem cells, and (iii) double-positive thymocytes derived from these pro-T cells after injection into Rag-deficient mice. The combined analyses of the different datasets in this unique experimental system highlighted the importance of both transcriptional and epigenetic repression in shaping the early phases of T-cell development.