Project description:MiRNAs play pivotal roles in regulating macrophage functions, including differentiation, polarization, recruitment, and activation of inflammation. in this review, to provide novel insight into macrophage differentiation-specific mitomiR signatures and associated mitochondrial pathways, we performed a global miRNA profiling on extracted mitochondria and total cell content of human macrophages. The THP-1 monocytic cell line was treated with phorbol-12-myristate-13-acetate (PMA) to induce differentiation into macrophages The human monocytic cell line THP-1 cells were differentiated into THP-1 derived macrophages with 40 ng/mL phorbol 12-myristate 13-acetate (PMA),The mitochondrial fraction of the cells were enriched and purified using anti-TOM22 microbeads (Miltenyi Biotec). After total RNA extraction, and using Taqman low-density arrays (TLDA), we performed the global miRNA profiling on extracted mitochondria and compared with total cell content
Project description:Allele-specific regulatory events are essential in many fundamental biological processes. In this study, we utilized the human monocytic cell line THP-1 and performed RNA immunoprecipitation sequencing (MeRIP-seq) to explore allele-specific regulation of N6-methyladenosine (m6A) RNA modifications. Recent research highlights the importance of allele-specific regulation of m6A (ASm6A) and the need for precise detection methods to elucidate its underlying mechanisms. To address limitations in current approaches, which often emphasize genetic variations rather than comprehensive regulatory mechanisms, we applied M6Allele, a meta-analysis framework based on hierarchical Bayesian models. This method enables accurate detection of ASm6A events at the peak level, providing new insights into allele-specific regulatory mechanisms of m6A modifications. Our findings from the THP-1 cell line contribute to a deeper understanding of ASm6A and its biological significance.
Project description:Eicosanoids are potent regulators of gene expression of inflammatory cells. Pro- (leukotrienes B4 and C4) and anti-indflammatory (lipoxins A4 and B4) eicosanoids have been described in the literature but the detailed impact of these lipid mediators on the gene expression pattern of monocytic cells has not been studied in detail. We cultured the permanent monocytic cell line MonoMac 6 for 12 h in the absence (solvent control) and presence of these eicosanoids and quantified the differential gene expression patterns using the microarray technology. Experiment Overall Design: MonoMac6 cells were grown according to the recommendations of the German Tissue and Cell Culture Collection (Braunschweig) to near confluency. Then 100 nM of the eicosanoids (leukotriene B4, leukotriene C4, lipoxin A4, lipoxin B4) were added to the culture medium as DMSO stock solution and the cells were further cultured for 12 h with the stimuli. Then the cells were harvested, washed and total RNA was extracted according to the Qiagen protocol. Total RNA was used for microarray hybridization.
Project description:Transcriptome analysis of the human monocytic cell line THP1 (ATCC TIB-202) treated or not with phorbol myristate acetate (PMA) +/- lipopolysaccharides-lipopolysaccharides binding protein (LPS-LBP)
Project description:We performed proteomics of mitochondria isolated from leukemia cell line. Digested peptides were labeled with TMT and separated in 9 fractions by High-pH fractionation Kit. Peptides were analyzed by SPS-MS3 mode.