Journal: bioRxiv

Article Title: The TET-dependent DNA demethylation pathway is the driving force of hematopoiesis

doi: 10.64898/2026.04.29.721744

Figure Lengend Snippet: The development of T cells is controlled by the TET dependent DNA demethylation pathway. A) Inactivation of TET enzymes results in the maintenance of high levels of methylation of genes essential for T cell development, according to DMRseq analysis (T_cells_KO - T cells isolated from the spleen of mice treated with tamoxifen; T_cells_C - T cells isolated from the spleen of control mice). B) PCA (B’), heatmap (B’’), and volcano plot (B’’’) indicate a significant difference in gene expression between TET-deficient (T_cell_KO) and control (T_cell_C) T cells. C) The heatmap shows that maintaining high levels of T cell gene methylation leads to a decrease in their expression. D) Many genes whose expression is reduced in TET-deficient T cells are necessary for their specification and maturation/function. At the same time, the expression of genes that are markers of HSPCs (Cd34, Cd117) is increased in TET-deficient T cells. Taken together, these results indicate the underdevelopment of these cells. CLP – common lymphoid progenitors, DN – double negative, DP – double positive

Article Snippet: Cd117 (cKit) positive mouse cells were isolated from bone marrow using CD117 MicroBeads according to the MACS protocol (130-091-224, Miltenyi Biotec, Auburn, CA USA).

Techniques: Methylation, Isolation, Control, Gene Expression, Expressing

Journal: bioRxiv

Article Title: The TET-dependent DNA demethylation pathway is the driving force of hematopoiesis

doi: 10.64898/2026.04.29.721744

Figure Lengend Snippet: Demethylation and activation of many hematopoietic genes are suppressed early in hematopoiesis in TET-deficient experimental mice. A) The TET dependent DNA demethylation pathway initiates the demethylation of hematopoietic genes as early as the myeloid/lymphoid progenitor stage. DMRs were identified by comparing the methylomes of experimental (Cd117_KO) and control (Cd117_C) Cd117 + cells using the DMRseq Bioconductor package. B) Changes in methylation patterns caused by inactivation of TET enzymes lead to significant changes in gene expression in Cd117 + cells as evidenced by PCA (B’), heatmap (B’’), and volcano plot (B’’’). C) The heatmap reflects changes in the expression of many genes involved in hematopoiesis in Cd117_KO vs Cd117_C cells. D) The panel shows the stages of hematopoiesis and changes in the expression of genes corresponding to these stages. A green downward arrow indicates decreased gene expression in TET-deficient Cd117 + cells, while a red upward arrow indicates increased gene expression. E) To examine the rate of Cd117 + cell proliferation, EdU was administered intraperitonially into experimental and control mice. The bone marrow of these animals was collected after 30 minutes and used for Cd117 + cell isolation. Cd117 + cells were labeled, and the percentage (%) of cells incorporating EdU was determined using flow cytometry (n=5, ****P-value < 0.0001). F) The percentage (%) of apoptotic (Annexin V+/PI-; Q4) and necrotic (PI+; Q1 and Q2) Cd117 + cells was determined using flow cytometry (n=5, ****P-value < 0.0001, *P-value < 0.05).

Article Snippet: Cd117 (cKit) positive mouse cells were isolated from bone marrow using CD117 MicroBeads according to the MACS protocol (130-091-224, Miltenyi Biotec, Auburn, CA USA).

Techniques: Activation Assay, Control, Methylation, Gene Expression, Expressing, Cell Isolation, Labeling, Flow Cytometry