dna methyltransferase inhibitor decitabine  (Selleck Chemicals)

Journal: bioRxiv

Article Title: DNA replication stress redistributes the histone methyltransferase G9a and activates lineage-specific gene expression

doi: 10.64898/2026.10.06.757090

Figure Lengend Snippet: (A) Dose-response curve of fetal hemoglobin induction (HbF, red, left axis) and cell viability (black, right axis) in HUDEP-2 cells treated with increasing concentrations of hydroxyurea (HU) (n=3). (B) Representative immunoblots of HUDEP-2 cells treated for 48 hr with the indicated doses of HU. Phosphorylation of CHEK1 at Ser345 (p-Chk1) is shown as a marker of DNA replication stress and phosphorylation of CHEK2 at Thr68 (p-Chk2) as a marker of DNA damage. Vinculin serves as a loading control. (C) Nascent RNA expression analysis of globin genes ( HBG1 , HBG2 , and HBB ) in HUDEP-2 cells at the indicated times following treatment with HU. Bar plots show the mean log2 fold change (log 2 FC) of primary transcript levels relative to vehicle at the indicated time points after treatment. Asterisks denote statistically significant (p < 0.00001) changes compared to vehicle (n=4). (D) Dose-response curve of HbF induction (red, left axis) and cell viability (black, right axis) in HUDEP-2 cells treated with increasing concentrations of aphidicolin, etoposide, decitabine, and methotrexate as indicated (n=3). (E) Immunoblot analysis of HUDEP-2 cells treated as described in (D) for p-Chk1, p-Chk2, and Vinculin. (F) Dose-response curves of HbF induction in primary human CD34 + hematopoietic stem and progenitor cells (HSPCs) treated with increasing concentrations of hydroxyurea or aphidicolin (n=3). (G) Single-cell RNA-seq analysis of CD34+ HSPCs treated with vehicle (Veh), HU, or aphidicolin (APH) from days 7–14 of erythroid differentiation and analyzed on day 14. (Left) UMAP projections showing density plots of cells from each treatment condition, overlaid on the reference hematopoietic differentiation landscape (gray). Clusters 1-3 are labeled. (Right) Dot plots showing HBG1 and HBG2 expression within erythroid clusters 1-3 for each treatment condition. Dot size represents the percentage of cells expressing each transcript and color indicates scaled, log-normalized average expression.

Article Snippet: The following drugs were used to induce replication stress: hydroxyurea (HU; Sigma-Aldrich, H8627 and EMD Millipore, 400046), aphidicolin (APH; Sigma-Aldrich, A0781), etoposide (Sigma-Aldrich, E1383), decitabine (DEC; MedChemExpress, HY-A0004), methotrexate (Sigma-Aldrich, A6770), camptothecin (Santa Cruz Biotechnology, SC-200871), gemcitabine (GEM; MedChemExpress, HY-17026), pomalidomide (POM; TCI Chemicals, P2074), and cytarabine (Selleck Chemicals, S1648).

Techniques: Western Blot, Phospho-proteomics, Marker, Control, RNA Expression, Analysis, Single Cell, RNA Sequencing, Labeling, Expressing

Journal: Nature Communications

Article Title: Histone deacetylase inhibition induces an antitumor IRF8-dependent trained immunity-like status in macrophages

doi: 10.1038/s41467-026-77904-3

Figure Lengend Snippet: a – c On d0, bone marrow (BM) cells were treated with epigenetic modifiers or vehicle (Con) in the presence of M-CSF to initiate macrophage differentiation. On d3, modifiers were washed out. Mature bone marrow-derived macrophages (BMDMs) were harvested on d6, and challenged with LPS or 4T1 conditioned medium (CM). A schematic diagram was created in BioRender. q, X. (2026) https://BioRender.com/d3iy1fx . Inflammatory cytokine gene expression ( b , qPCR) and protein secretion ( c , ELISA) from these BMDMs were quantified ( n = 3 samples). d , Heatmap displaying relative chemokine secretion levels from in vitro Vor-primed or Con macrophages ( n = 4 samples). e Experimental schematic for monocyte adoptive transfer. (Top) E0771 tumor-bearing mice were treated with Vor or Con (d7-d17), monocytes were sorted on d17. (Bottom) Macrophage-depleted Lyz2 Cre-ERT2 Csf1r fl/fl mice bearing E0771 tumors were reconstituted with Con-Mono or Vor-Mono. Representative images ( f ) and growth curves ( g ) of E0771 tumors ( n = 5 mice). Flow cytometric analysis of TAMs and CD8 + T cells. h CXCL9, CCL8, and MHC-II expressions on macrophages, CX3CR1 mid MHC-II high subset proportion within macrophages. i CD8 + T-cell frequency (% of CD3 + ) and proportions of GZMB + and Perforin + cells (% of CD8 + ) ( n = 5 samples). j Treatment in MMTV-PyMT mice. PBS or Vor (q.o.d.) from 12 weeks; at 14 weeks, TAMs were sorted and co-cultured with 4T1/E0771 for phagocytosis and cytotoxicity assays. k Representative images of mammary tumors from MMTV-PyMT mice ( n = 5 mice). l Quantification of phagocytosis and cytotoxicity against 4T1 cells by TAMs ( n = 10 samples). m Schematic of macrophage adoptive transfer. (Top) BM cells were treated with Con or Vor (d0-d3); mature BMDMs were harvested on d6. (Bottom) Macrophage-depleted Lyz2 Cre-ERT2 Csf1r fl/fl mice bearing E0771 tumors were reconstituted with Con- or Vor-Macrophages. n Growth curves of subcutaneous E0771 tumors (from Fig. 2m) receiving Con-Macro versus Vor-Macro ( n = 5 mice). o Frequencies of intratumoral MHC-II + macrophages, Granzyme B (GZMB) + CD8⁺ T cells, and PD-1 + TIM-3 + CD8⁺ T cells ( n = 5 samples). Schematic diagrams ( e , j , m ) were created with BioGDP.com. Data are presented as mean ± SD, by one-way ANOVA followed by Dunnett’s multiple comparisons test ( b , c ), two-way ANOVA ( g , n ) or two-sided Mann-Whitney U test ( h , i , l , o ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Source data and exact P -values are provided as a Source Data file.

Article Snippet: The epigenetic modifiers (all purchased from MedChemExpress) included the DNA methylation inhibitors decitabine (10 nM) and azacitidine (1 μM); the histone methylation modulators tazemetostat (4 μM), EPZ004777 (1 μM), GSK-J4 (10 μM), and seclidemstat (1 μM); the histone acetylation modulators vorinostat (10 nM), chidamide (10 nM), and selisistat (0.5 μM); and the epigenetic reader inhibitor JQ1 (2 μM).

Techniques: Derivative Assay, Gene Expression, Enzyme-linked Immunosorbent Assay, In Vitro, Adoptive Transfer Assay, Analysis, Cell Culture, MANN-WHITNEY, Real-time Polymerase Chain Reaction