dnmt1 inhibitors ezh2 inhibitor tazemetostat epz 6438 selleckchem s7128 dnmt1 inhibitor  (Selleck Chemicals)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Novel feeder-cell-free 3C system promotes the transition from primed to formative pluripotency, self-renewal, and germline differentiation in rabbit embryonic stem cells in vitro

doi: 10.3389/fcell.2026.1841436

Figure Lengend Snippet: Transcriptome analysis and gene expression. (A) Volcano plot of gene expression in the 3C and iFLY systems. Blue dots mark significantly downregulated genes; red dots mark upregulated genes. Naïve pluripotency markers (e.g., ZFP42 ) and formative pluripotency markers (e.g., OTX2 and LIN28B ) were upregulated in the 3C system, while primed pluripotency markers (e.g., NODAL , HOXA1 , and TBXT ) were downregulated. (B) KEGG analysis showed significant upregulation of the Ras and MAPK pathways in 3C system-derived rbES cells. The upregulated genes were primarily enriched in pathways related to cell communication and adhesion, including cell adhesion molecules, ligand-receptor interaction, Rap1 signaling, and MAPK signaling, suggesting enhanced cell-cell interaction and environmental responsiveness in the 3C state. (C) KEGG analysis revealed significant downregulation of the PI3K-Akt and Wnt pathways in 3C system-derived rbES cells. (D) RT-qPCR showed that in the 3C system, POU5F1 , NANOG , ZFP42 , KLF17 , LIN28B , and OTX2 were significantly upregulated, while NODAL and TBXT were significantly downregulated compared with their levels in cells from the iFLY system. β-ACTIN was used to normalize the data, which are presented as the mean ± SD. Independent samples t-tests were used for statistical analysis. **, ***, and **** indicate P < 0.01, <0.001 and <0.0001, respectively. (E) Schematic illustrating the establishment of the feeder-free 3C culture system for rbES cells.

Article Snippet: Using a feeder-free culture system, we evaluated the effects of several cell factors at different concentrations on the pluripotency of rbES cells, including IWP2, a Wnt/β-catenin inhibitor (cat. no. T2702, TargetMol), at concentrations of 1, 5, and 10 μM; PD0325901, a MEK inhibitor (cat. no. T6189, TargetMol), at concentrations of 0.2, 1, and 3 μM; CHIR99021, a GSK-3β inhibitor (cat. no. T2310, TargetMol), at concentrations of 0.2, 1, and 3 μM; DZNep, an EZH2 methyltransferase inhibitor (cat. no. T6360, TargetMol), at concentrations of 10, 50, and 100 nM; and TSA, an HDAC inhibitor (cat. no. T6270, TargetMol), at concentrations of 5, 20, and 50 nM.

Techniques: Analysis, Gene Expression, Derivative Assay, Quantitative RT-PCR, Significance Assay, Protein-Protein interactions, RNA Sequencing, Real-time Polymerase Chain Reaction