Project description:metabolite levels provided by UM platform (Creative Dynamics Inc, NY, USA) (the data is raw abundance. Mapping was applied on log10 transformed data)
Project description:The transcriptional and metabolic profiles of rosettes of three independent Arabidopsis thaliana ecotype Wassiliewska lines overexpressing the soybean early noduline GmENOD40 gene were compared to those of wt, and a biomarker analysis was performed. ENOD40 overexpression resulted in 382 modulated genes, 142 of which upregulated and 165 downregulated in all the three transformed lines. Cell wall, membrane, hydrolase activity on glycosidic bonds, defense response and response to stimulus gene categories were overrepresented in the modulated genes compared to TAIR complete database. Metabolomics analysis (through liquid chromatography-mass spectrometry) allowed to putatively identify 13 different glucosinolates, some flavonoids, sinapic and ferulic acid derivatives, hydroxybenzoic acid derivatives, non aromatic organic acids, amino acids. Positive genes biomarkers of the transformed plants were mainly involved in cell wall turnover, whereas positive metabolite biomarkers were the methylthioalkyl-glucosinolates. The transcriptional profile of rosettes of three independent Arabidopsis thaliana ecotype Wassiliewska lines overexpressing the soybean early noduline GmENOD40 gene were compared to those of wt, and a biomarker analysis was performed.
Project description:The transcriptional and metabolic profiles of rosettes of three independent Arabidopsis thaliana ecotype Wassiliewska lines overexpressing the soybean early noduline GmENOD40 gene were compared to those of wt, and a biomarker analysis was performed. ENOD40 overexpression resulted in 382 modulated genes, 142 of which upregulated and 165 downregulated in all the three transformed lines. Cell wall, membrane, hydrolase activity on glycosidic bonds, defense response and response to stimulus gene categories were overrepresented in the modulated genes compared to TAIR complete database. Metabolomics analysis (through liquid chromatography-mass spectrometry) allowed to putatively identify 13 different glucosinolates, some flavonoids, sinapic and ferulic acid derivatives, hydroxybenzoic acid derivatives, non aromatic organic acids, amino acids. Positive genes biomarkers of the transformed plants were mainly involved in cell wall turnover, whereas positive metabolite biomarkers were the methylthioalkyl-glucosinolates.
Project description:Comprehensive and quantitative information of the thermophile proteome is an important source for understanding of the survival mechanism under high growth temperature. Thermoanaerobacter tengcongensis (T. tengcongensis), a typical anaerobic thermophilic eubacterium, was selected to quantitatively evaluate its protein abundance changes in response to four different temperatures. Thermoanaerobacter tengcongensis proteins were trypsine digested, separated with high-pH RP, and identified with MS/MS analysis. The raw MS/MS data were converted into MGF format by Proteome Discoverer 1.2 (Thermo Fisher Scientific, Waltham, MA, USA). And the exported MGF files were searched by Mascot 2.3.02 (Matrix Science, Boston, MA, USA) against the database with all 2588 predicted proteins in T. tengcongensis downloaded from NCBI (NCBI reference sequence: NC_003869.1). An automatic decoy database search was performed. Several parameters in Mascot were set for peptide searching, tolerance of one missed cleavage of trypsin, Carbamidomethyl (C),iTRAQ8plex (K) and iTRAQ8plex (N-term) as fixed modification, iTRAQ8plex (Y),Oxidation (M) as variable modification. The precursor mass tolerance was 10 ppm, and the product ion tolerance was 0.02 Da.
Project description:These RNAseq data are from germinating cyp79B2, cyp79B3, myb28, myb29 (qko), cyp79B2, cyp79B3 (cyp) and myb28 myb29 (myb) mutant seeds. Col-0 genotype was used as the Arabidopsis' WT reference ecotype and all mutant seeds are from the Col-0 genetic background. Samples were sown on agarose 0,8%, supplemented or not by nitrate KNO3 10Mm or potassium thiocyanate KSCN 8µM. RNA isolation was performed using NucleoSpin® RNA Plant and Fungi kit. RNA quantification was obtained with a NanoDrop ND-100 (NanoDrop Technologies, DE, USA) and the RNA Integrity was measured with a 4100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). cDNA library construction and sequencing were performed with DNBseq™ technology at Beijing Genomics Institute BGI. FastQC was used to check read quality, mapping analysis and transcript quantiifcation were performed using a quasi-mapping alignment from Salmon.
Project description:We used an in vivo short hairpin RNA (shRNA) screening approach to identify genes that are essential for MLL-AF9 acute myeloid leukemia (AML). We found that Integrin Beta 3 (Itgb3) is essential for murine leukemia cells in vivo, and for human leukemia cells in xenotransplantation studies. In leukemia cells, Itgb3 knockdown impaired homing, downregulated LSC transcriptional programs, and induced differentiation via the intracellular kinase, Syk. In contrast, loss of Itgb3 in normal HSPCs did not affect engraftment, reconstitution, or differentiation. Finally, we confirmed that Itgb3 is dispensable for normal hematopoiesis and required for leukemogenesis using an Itgb3 knockout mouse model. Our results establish the significance of the Itgb3 signaling pathway as a potential therapeutic target in AML. R940406 (R406, the active metabolite of fostamatinib) was supplied by Rigel Pharmaceuticals, Inc., South San Francisco, CA, and AstraZeneca Pharmaceuticals, Wilmington, DE, USA. R406 was resuspended in dimethyl sulfoxide (DMSO) (Sigma-Aldrich) and stored at −80°C. . HL-60, U937 and KG-1 cell lines were purchased from the American Type Culture Collection. MOLM-14 cell lines were provided by Dr. Scott Amstrong (Dana-Farber Cancer Institute, Boston MA, USA.) All cell lines were maintained in RPMI 1640 (Cellgro) supplemented with 1% penicillin-streptomycin and 10% fetal bovine serum (FBS, Sigma-Aldrich) at 37 °C with 5% CO2.
Project description:Metabolic reprogramming in cancer and immune cells occurs to support their increasing energy needs in biological tissues. Here we propose Single Cell SPAtially resolved METabolic (scSpaMet) framework for joint protein-metabolite profiling of single immune and cancer cells in male human tissues by incorporating untargeted spatial metabolomics and targeted multiplexed protein imaging in a single pipeline. We utilized the scSpaMet pipeline to profile cell types and spatial metabolomic maps of 19507, 31156, and 8215 single cells in human lung cancer, tonsil and endometrium tissues, respectively. ScSpaMet analysis revealed cell type-dependent metabolite profiles and local metabolite competition of neighboring single cells in human tissues. Deep learning-based joint embedding revealed unique metabolite states within cell types. Trajectory inference showed metabolic patterns along cell differentiation paths. Here we show scSpaMet’s ability to quantify and visualize the cell-type specific and spatially resolved metabolic-protein mapping as an emerging tool for systems-level understanding of tissue biology.
Project description:In this study, integrated transcriptomics, proteomics and metabolomics approaches were applied to investigate the molecular responses of O3 in the leaves of two-weeks old rice (cv. Nipponbare) seedlings exposed to 0.2 ppm O3 for a period of 24 h. Based on the morphological alteration of O3-exposed rice leaves, transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that O3 triggers a chain reaction of altered gene, protein and metabolite expressions involved in multiple cellular processes in rice. Also investigated were the molecular responses in the leaves of two-weeks old rice (cv. Nipponbare) seedlings under continuous light and pure air (as a positive control for ozone exposure experiments) for a period of 24 h. Transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that continuous light and growth for 24 h also triggers a chain reaction of altered gene expressions involved in multiple cellular processes in rice, but different from those against ozone, in general. Keywords: Ozone fumigation response
Project description:The c-MET signaling axis is increasingly implicated in tumorigenesis and chemioresistance. In this study, we investigated gene expression modifications induced by SU11274 (Selleck Chemicals, Boston, USA), a novel selective c-MET inhibitor, in a pair of isogenic multiple myeloma (MM) cell lines either sensitive (RPMI8226) or multi-resistant, highly c-MET expressing (RPMI8226/R5) cells. On the whole, RPMI8226/R5 cells after SU11274 were characterised by wider and diverse gene expression modifications than RPMI8226, indicating that c-MET over-expression, and its inhibition, is an important aspect of the adaptive response associated to drug resistance.