Project description:Motor unit remodelling involving repeated denervation and re-innervation occurs throughout life. The efficiency of this process declines with age contributing to neuromuscular deficits. We investigated differentially expressed genes (DEG) in muscle following peroneal nerve crush to model of motor unit remodelling in C57Bl6 mice. Muscle RNA was isolated at 3 days post-crush, RNA libraries were generated using poly-A selection, sequenced.
Project description:The RNA sequencing experiment is part of the study: “Modulating Redox Balance Restores Azacytidine Efficacy in Hypomethylating Agent Resistant Disease.” In the study we generated myelodysplastic syndrome/acute myeloid leukemia (MDS/AML) OCI-M2 cell line that is resistant to hypomethylating therapy by 5-azacytidine (AZA). By modulation of the redox environment via modification of redox sensor KEAP1 using sulforaphane (SFN) in these cells we were able to restore sensitivity to AZA. We used RNA sequencing to define transcriptomic differences between AZA sensitive (AZA-S) and AZA resistant (AZA-R) cells and to characterize how the transcriptome is changing upon treatment of these cells with AZA, SFN and combination of both.
Project description:The changes in the transcriptome of leaves between 0, 9, 13 and 17 days of cold acclimation were analyzed using RNA-Seq in Veyo and Falster genotypes of perennial ryegrass.
Project description:DNA double-strand breaks (DSBs) are toxic to mammalian cells. However, during meiosis, more than 200 DSBs are generated deliberately, to ensure reciprocal recombination and orderly segregation of homologous chromosomes. If left unrepaired, meiotic DSBs can cause aneuploidy in gametes and compromise viability in offspring. Oocytes in which DSBs persist are therefore eliminated by the DNA-damage checkpoint. The checkpoint’s downstream effectors that trigger oocyte death, thereby preserving genome stability across the generations, are unknown. Here we show that the DNA-damage checkpoint eliminates oocytes via the pro-apoptotic BCL-2 pathway members Puma, Noxa and Bax. Deletion of these factors prevents oocyte elimination in recombination-repair mutants, even when the abundance of unresolved DSBs is high. Remarkably, surviving oocytes can extrude a polar body and be fertilised, despite chaotic chromosome segregation at the first meiotic division. Our findings raise the possibility that allelic variants of the BCL-2 pathway could influence the risk of embryonic aneuploidy.
Project description:Human valvular endothelial cells were co-cultured with THP-1 monocytes in high glucose (33mM) or normal glucose (5mM) levels for 2 hours. Afterwards cells were lysed and total cellular RNA was extracted using TRIzol reagent.
Project description:Metabolic dysfunction-associated fatty liver disease is the most prevalent liver disease and affects a quarter of the global population. Estrogens are associated to safeguard the liver from metabolic diseases. We fed male and female mice a control or high-fat diet for 13 weeks. Only male mice developed fatty livers. We injected a subset of male mice fed a high-fat diet with four different estrogen receptor (ER) agonists for the last three weeks of the high-fat diet, activating the nuclear ERalpha and ERbeta. Livers were collected and flash-frozen before RNA isolation, DNAse treatment, library preparation and sequencing on an Illumina NextSeq 500 instrument.
Project description:In this experiment, SUM149 IBC cells were cultured using baseline medium (N=3), and in baseline medium supplemented with increasing concentration of Lapatinib to induce resistance (N=3; rSUM149). Resistant reversal SUM149 cells (rrSUM149) were then generated by culturing in rSUM149 cells in baseline medium. Gene expression data were generated to identify differentially expressed genes, as well as differentially activated proteins and pathways during Lapatinib resistance and resistance reversal. Gene expression were also integrated with proteome expression data generated under the same conditions.
Project description:This experiment aims to determine the genes that respond to protoplasting treatment in a time course of Arabidopsis seed germination, so that they can be filtered out from single-cell RNA-seq analysis which involves protoplasting.
Project description:The aim of this experiment is to identify genes in the bacteriophage S-PM2d that are deferentially expressed in response to light intensity during infection. We were interested in light since the host (Synechococcus) obtains all energy and carbon for growth from light, through photosynthesis. The amount of light entering the cell is therefore approximately proportional to the amount of energy that can be generated. Therefore we wanted to increase the amount of energy available for an infecting virus (S-PM2) and observe the physiological and transcriptional response of the virus. This is particularly interesting because the virus itself encodes genes involved in harvesting this light into energy.
Project description:The microRNA miR-96 is important for hearing; mutations in the seed region result in dominant progressive hearing loss in mice and humans. Mir96 is expressed in the sensory hair cells of the organ of Corti along with Mir182 and Mir183. miR-96 is a master regulator of hair cell development, controlling many genes in the organ of Corti, but the role of miR-182 and miR-183 in the hair cells is unknown. We carried out RNA-seq on mice carrying a knockout allele of Mir182, and mice carrying a double knockout allele of Mir183 and Mir96 (Mir183/96). RNA was extracted from the organ of Corti from P4 homozygotes and sex-matched wildtype littermates. Strand-specific libraries were prepared using the NuGEN Ovation Mouse RNA-Seq System 1-16 kit and sequenced on an Illumina HiSeq 2500 machine as paired-end 125bp reads.