Project description:This project aimed to identify novel endosomal proteins and therefore cellular fractionation experiments were performed. Adherent mouse embryonic fibroblast were scraped on ice into hypotonic lysis buffer (10 mM HEPES-KOH pH 7.2, 0.25 M sucrose, 1 mM EDTA, 1 mM MgOAc and protease & phosphatase inhibitors (PhosSTOP and Complete mini tablets from Roche). Cell membranes were fragmented with French Press and nuclei removed with 10 min 1000 x g centrifugation. Plasma membrane fraction was collected with 10 min 10 000 x g centrifugation and endosomal/cytoplasmic fractions with 1h 100,000 x g centrifugation. Membrane fractions were washed at least once with lysis buffer and cytoplasmic fractions were centrifuged twice. All fractionation steps were performed at +4°C or on ice. All fractions were dissolved in sample buffer for immunoblotting
Project description:Gene expression profiling of pre-adipocytes 3T3-L1 reveals anti-adipogenic potential to metabolic associated diseases through whole transcriptomic analysis. We evaluated the effects of Tsuruazuki extract on pre-adipocytes 3T3-L1. We performed an untargeted whole-genome transcriptome analysis to explore functionality of Tsuru on 3T3-L1 cells.
Project description:We sought to determine the transcriptomic impacts of artemisinin, Artemisia annua extract, and Artemisia afra extract on M. tuberculosis. Log phase cultures were treated with lethal doses for four hours or with inhibitory or sub-inhibitory doses for 24 hours. RNA was collected from untreated controls at the same timepoint.
Project description:Gene expression profiling reveals mesoderm lineage-specific genes were more significantly regulated with the most prominent regulated DEGs were identified in adipose tissue. We evaluated the effects of Tsuruazuki extract on human amniotic epithelial stem cells. We performed an untargeted whole-genome transcriptome analysis to explore functionality of Tsuru in a stem cell-based tool.
Project description:Impact of French Maritime Pine Bark Extract and Methylphenidate on the Gut Microbiota of paediatric Attention-Deficit Hyperactivity Disorder Patients
Project description:Euphorbia pithyusa data were performed on Q-TOF agilent instrument. Extract of aerial parts of plant and the chromatographic fractions were analyzed in untargeted LC-MS/MS mode.
Project description:Procedure details are reported in McCafferty et al. eLife2022;11e81977. Briefly cilia extract from Tetrahymena thermophila SB715 was fractionated on a mixed bed IEX HPLC column and all resulting fractions were crosslinked by addition of DSSO to 0.5mM. Following reduction, alkylation, trypsin digestion, and desalting, mass spectra were collected on an Orbitrap Fusion Lumos using a DDA MS2-MS3 method and spectra were processed in ProteomeDiscoverer 2.3 using the Xlinkx node to identify crosslinks. The look-up database for crosslink identification was generated from the same .RAW files using a standard Basic workflow to identify proteins present in the fractions.