Project description:The differentiation of macrophages from monocytes is a tightly controlled and complex biological process. Although numerous studies have been conducted using biochemical approaches or global gene/protein profiling, the mechanisms of the early stages of differentiation remain unclear. Here we used THP1 cells to model the differentiation and studied their phosphoproteome perturbation during the early differentiation process
Project description:The nuclear proteome harbors a sheer number of regulatory proteins, including transcription factors (TFs). Profiling of nuclear proteome during all-trans-retinoid acid(ATRA)-induced differentiation of HL60 cells allows to unveil molecular mechanisms of granulocytic maturation. It is especially important to have an understanding of molecular perturbations at the early stages of the differentiation process. Applying proteomic profiling using isobaric labeling coupled with alkaline fractionation (TMT/2D) we identified 1860 nuclear proteins with high confidence (FDR<0.01, at least 2 unique peptides per protein). Among them 136, 226, 280, 312 and 241 proteins were found to be altered at 3, 6, 9, 12, and 72 h in HL60 cell nuclear fraction under ATRA treatment.
Project description:The purpose of this study was to determine the miRNA expression profile of in vitro differentiation of human skeletal muscle cells and to couple changes in individual miRNA expression to effects on target genes by transcriptome profiling of mRNA expression. mRNA expression profiling at three different time points during the in vitro differentiation process of human skeletal muscle cells from three subjects. RNA was harvested from myoblasts before and 4 and 10 days after induction of differentiation. Temporal, time-course design with paired analysis (3 subjects). Biological replicates: 3 at day 0, 3 at day 4, and 3 at day 10. One replicate per array.
Project description:Assessment of genomewide nascent transcription activity and transcriptomic profiles across different stages of directed differentiation from human pluripotent stem cells (hPSC) into small intesitnal organaoids
Project description:The mechanisms of neural differentiation of human induced pluripotent stem cells (hiPSCs) have been widely investigated, however the genes that play key regulatory role, especially involved in mitochondrial function are still unidentified. In this report transcriptome analysis of hiPSC differentiating into neural stem cells (NSC), early neural progenitors (eNP) and neural progenitors (NP) were performed. Gene Ontology and Network Analysis was implemented considering especially mitochondrial genes at all developmental stages and upon stimulation by idebenon (IDB) and bezafibrate (BZ) at NP stage. The genes strongly up-regulated during the process of differentiation in all tested stages belong mainly to: WNT; Cadherin and Gonadotropin-releasing hormone receptor signaling pathways. The interaction network shows that the transcription factors including HOXD3, ASCL1, HES5, and ZEB1 are possible key players in this process. Mitochondrial unfolded protein response (UPRmt), ROS defense, and pyruvate metabolism were the strongest up-regulated mitochondrial processes in all analyzed stages of neural differentiation. IDB and BZ treatment at NP stage induced different set of nuclear encoded mitochondrial genes involved in respiratory electron transport chain and fatty acid metabolic process respectively. Thus PARP agonist-mediated stimulation of mitochondrial genes influence the process of neural differentiation despite of mitochondrial biogenesis, but through the specific receptor pathways.
Project description:During eukaryotic transcription, RNA polymerase II undergoes dynamic post-translational modification on the C-terminal domain (CTD) of the largest subunit , generating a sophisticated PTM landscape for the spatiotemporal recruitment to transcriptional regulators. To delineate the protein interactomes recruited to Pol II at different stages of transcription, we in vitro reconstructed phosphorylation patterns of the CTD at Ser5 and Ser2 positions, the hallmark phosphorylation at the initation and productive elongation stages of transcription, respectively. Distinctive protein interactomes indicates different proteins are recruited to RNA polymerase II at different stages of transcription by the phosphorylation of Ser2 and Ser5 of the CTD heptads. Calcium Homeostasis Endoplasmic Reticulum Protein (CHERP) specifically binds to the Ser2 of the heptad. The loss of the interaction between CHERP and Pol II results in broad alternative splicing events. Our method points to a new method to distinguish the PTM codes that coordinate the transcription process.
Project description:Generation of oligodendrocytes (OLs) is a sophisticated multistep process, mechanistic underpinnings of which are not fully understood and demand further investigation. To systematically profile proteome dynamics during human embryonic stem cell (hESC) differentiation into OLs, we applied in-depth quantitative proteomics at different developmental stages and monitored changes in protein abundance using a multiplexed tandem mass tag (TMT) based proteomics approach. Findings: Our proteome data provided a comprehensive protein expression profile that highlighted specific expression clusters based on the protein abundances over the course of human OL lineage differentiation. The proteome profile of OL lineage cells revealed 378 proteins that were specifically up-regulated only in one differentiation stage. In addition, comparative pairwise analysis of differentiation stages demonstrated that abundances of 352 proteins differentially changed between consecutive differentiation time points. Our results highlighted the eminence of the planar cell polarity (PCP) signaling and autophagy (particularly macroautophagy) in the progression of OL lineage differentiation