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: Apoptotic and necrotic cells were detected using the Annexin V-FITC Kit (130-092-052, Miltenyi Biotec, Auburn, CA USA) and the LSR-Fortessa-HTS instrument.

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

Journal: Blood Neoplasia

Article Title: Targeting the G-protein–coupled estrogen receptor: a novel therapeutic strategy in cutaneous T-cell lymphoma ∗

doi: 10.1016/j.bneo.2026.100213

Figure Lengend Snippet: G-1 induces apoptosis in CTCL cells. (A) For a 48-hour period, CTCL cells were exposed to either 500nM G-1 or DMSO. Cells were then stained for 20 minutes with Annexin V-FITC and 7-AAD. Annexin V–positive cells were analyzed by flow cytometry. Representative data shown from n = 3 independent experiments (left panel) and mean value of n = 3 independent experiments with SD (right panel). (B) The activation of caspase-3/7 by G-1 was assessed using the Caspase-Glo 3/7 assay. Cells were treated for 24 hours with 500nM G-1. The resulting caspase activity was normalized against cells treated with DMSO, which served as the untreated control. Mean values of n = 3 independent experiments with SD are plotted. (C) Different durations of treatment with DMSO or 500nM G-1 were applied to HuT and MyLa cells. Western blot analysis was used to evaluate protein expression levels. GAPDH served as a loading control. Representative plot of n = 3 independent experiments. Western blots were imaged digitally. Brightness and contrast were adjusted globally for each blot to improve clarity; no individual lanes were modified or removed. (D) Densitometric quantification of western blots shown in panel C. Band intensities were quantified using Image Studio, normalized to the loading control and expressed as fold change relative to DMSO (set to 1). Bars represent mean ± SD of n = 3 independent experiments. 7-AAD, 7-aminoactinomycin D; cIAP1, cellular inhibitor of apoptosis protein-1; DMSO, dimethyl sulfoxide; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PARP, poly(ADP-ribose) polymerase; SD, standard deviation.

Article Snippet: After treatment, cells were collected, resuspended in Annexin V binding buffer, and incubated with Annexin V fluorescein isothiocyanate and 7-aminoactinomycin D (7-AAD; Miltenyi Biotech, Bergisch Gladbach, Germany) for 20 minutes at room temperature, protected from light.

Techniques: Staining, Flow Cytometry, Activation Assay, Caspase-Glo Assay, Activity Assay, Control, Western Blot, Expressing, Modification, Standard Deviation