Project description:To model and characterize the chromatin regulatory landscape in Neural stem cell before and after Phf2 knockout, we performed HiC to map large-scale 3D architectural rewiring in WT and Phf2-KO NSC.
Project description:The establishment of C2C12 cell line with knockout of TAOK1 expression using CRISPR/Cas9 system. C2C12 cells with TAOK1 knockout were designated as C2C12-TAOK1-KO, while those transfected with an empty vector served as the control group (C2C12-Mock), non-transfected cells served as the blank group (C2C12-Blank). The C2C12-Blank, C2C12-Mock and C2C12-TAOK1-KO cells (10000 cells/mL) were seeded into 100 mm cell culture dishes and induced to differentiation into myotubes. Then, cells were obtained by trypsin digestion and centrifugation. Samples from three independent experiments were collected.
Project description:PHF2 demethylates lysine 9 in histone 3 and acts as a tumor suppressor. In liver pathogenesis, PHF2 has dual effects. PHF2 induces simple hepato-steatosis by epigenetically coactivating ChREBP. When lipid accumulation goes serious, it protects the liver from fibrogenesis by facilitating Nrf2 activity, a major transcription factor in the defense against oxidative stress during non-alcoholic fatty liver disease. However, little is known regarding the functions of PHF2 in liver cancer cells. Therefore, to investigate the possible roles of PHF2 and find new interacting proteins, we performed an LC-MS/MS assay combined with an immunoprecipitation assay.
Project description:The GFP or Phf2 were overexpressed in the liver of C57Bl6/J mice through adenoviral gene delivery. Mice were studied in the fed state 3 weeks later. We used microarray analysis to detail the global programme of gene expression underlying Phf2 overexpression and identified distinct classes of up and down regulated genes.
Project description:Long-term memory formation is attributed to experience-dependent gene expression. Dynamic changes in histone methylation status are essential for epigenetic regulation of memory consolidation-related genes. Here, we demonstrated that plant homeodomain finger protein 2 (PHF2) histone demethylase is upregulated in the mouse hippocampus during experience and plays an essential role in memory formation. QuantSeq analysis was performed to examine differential gene expression in the hippocampus of WT and PHF2 t/g mice. Transgenic mice were created by injecting the CMV-Flag-PHF2 vector into fertilized eggs from C57BL6 mice. Transgenic lines were established from 9 founders that were identified via PCR-based genotyping. Among the transgenic lines, mice with high PHF2 expressed brains were singled out for breeding. First, selected heterozygote males were bred with heterozygote females which produced homozygote, heterozygote and wild littermates. Since it is difficult to differentiate between hetero and homo mice with genotyping, we separated homo mice from hetero mice by breeding each of hetero and homo mice with wild type mice and confirming the genotypes of their offspring. In other words, hetero mice bred with wild type mice would have both wild type and hetero genotype of offspring, while homo mice bred with wild type mice would only have hetero genotype of offspring. Through this process, we particularly selected the homo mice and this transgenic line was used and maintained for this experiment.
Project description:Histone H3 lysine 9 methylation (H3K9me) is essential for cellular homeostasis; however, its contribution to development is not well established. Here, we demonstrate that the H3K9me2 demethylase PHF2 is essential for neural progenitor proliferation in vitro and for early neurogenesis in the chicken spinal cord. Using genome-wide analyses and biochemical assays we show that PHF2 controls the expression of critical cell cycle progression genes, particularly those related to DNA replication, by keeping low levels of H3K9me3 at promoters. Accordingly, PHF2 depletion induces R-loop accumulation that leads to extensive DNA damage and cell cycle arrest. These data reveal a role of PHF2 as a guarantor of genome stability that allows proper expansion of neural progenitors during development.
Project description:Important hallmark of neurogenesis is activation of quiescent neural stem cells (NSC) in dentate gyrus. Precise mechanisms of NSC activation are not fully clear. In the current study we interrogate the role of homeodomain finger protein 2 (Phf2) in this process. Here we performed mouse transcriptome RNA-seq profiling of epigenetic regulator Phf2 in neural stem cells without and with knock-down in order to uncover mechanistic components/pathways affecting protein translational capacity in activated NSC.
Project description:The precise regulation of neural stem cell quiescence and activation is crucial for normal brain development and lifelong nwurogenesis. However, the mechanisms that control neural stem cell activation remains poorly understood. In this study, we investigate the role of Phf2 in this process.