Project description:3 samples of R1, R2 and R3 bone marrow monocytes were compared from 3 biological replicates in 3 separate experiments. R1, R2 and R3 were sorted from triplicate experiments from pools of mice
Project description:Male and female collagen-enriched fraction of cartilage from bovine knees. Full thickness cartilage plugs without subchondral bone were digested with chondroitinase for 24 hours to remove proteoglycans. Cartilage was further decellularized with SDS.
LUM1_811145: channels 126, 127, 128 female, channels 129, 130, 131 male
LUM1_833734: channels 126, 127, 128 female, channels 129, 130, 131 male
Project description:The mouse liver mitochondrial proteome was analysed in four different mouse groups (allocation of the samples to Exp1 and Exp2 in brackets): AL (ad libitum fed) C57Bl6 (Exp1 129, Exp1 130, Exp2 130), DR (dietary restriction) C57Bl6 (Exp1 126, Exp1 127, Exp1 128), AL ICRFa (Exp2 128, Exp2 129) and old ICRFa (Exp2 126, Exp2 127). To achieve quantitative standardisation between Exp1 and Exp2, all 10 samples were pooled and one portion of the pool was anlaysed in each Exp (Exp1 131 and Exp2 131). Following tryptic digest and TMT 6-plex labelling, samples were separated by OffGel electrophoresis and all fractions were subsequently analysed by LCMSMS on an Orbitrap LTQ XL. One survey scan (res. 30,000) was followed by a high energy HCD scan (res. 6000) and a low energy CID scan in the LTQ. HCD data were used for the quantitation and CID data for the identification of peptides. Data were searched using Mascot (v. 2.2) through the Proteome Discoverer interface. Peptide raw quantitations were extracted as text files and further processed using dpeaqms (http://r-forge.r-project.org/projects/dpeaqms/) to obtain probability values for the differences in protein amounts for specific proteins between the different animal groups.
Project description:Arabidopsis thaliana mutant sr45-1 has an altered flower shape. sr45 is a splicing regulator. In this study, we examined the proteins from inflorescence of sr45-1 mutant plants and wild-type. Wild type TMT labels: 126, 128, 130. sr45-1 TMT labels: 127, 129, 131.
Project description:Samples 1012398_F1-F8: Using recombinant purified mCherry-tagged FUS-IDR we reconstituted condensates in a soluble nuclear extract isolated from U2OS cells. The extract was fractionated by centrifugation into a supernatant and pellet. Labels 126, 127, 128 are supernatant replicates and labels 129, 130, 131 are pellet replicates.
Samples 994350_F1-8: Using recombinant purified mEGFP-tagged MED1-IDR we reconstituted condensates in a soluble nuclear extract isolated from U2OS cells. The extract was fractionated by centrifugation into a supernatant and pellet. Labels 126, 127, 128 are supernatant replicates and labels 129, 130, 131 are pellet replicates.
For the proximity biotinylation method, we used SILAC (Lys-6, Arg-6) in the media of cells expressing the control while both FUS-IDR and MED1-IDR cells were grown in natural isotope conditions. Cells were subject to the biotinylation protocol followed by nuclear extract preparation. Light and heavy extracts were mixed at a 1:1 ratio followed by streptavidin pulldown, stringent washing, and elution. Two independent replicates of SILAC data were collected for both FUS-IDR (QEX2_1011001 and ECL1_1073837) and MED1-IDR (QEX2_996337 and ECL1_1078014).
Project description:The samples are in vitro cultured oligodendrocyte precursor cells in differentiation medium for 3 days. There are 2 groups: D3HET (ATG5F/+ CNP-Cre, TMT channels 126, 127, 128) and D3KO (ATG5F/F; CNP-Cre, TMT channels 129, 130, 131), with each group comprising 3 biological replicates. Subsequently, all the samples were lysed and subjected to TMT (Tandem Mass Tag) labeling, high pH fractionation into 8 fractions (F1-F8).
Project description:We reconstituted arrays of CTCF binding sites (L1, L2, L3, L4, R1, R2 and R3) and devised a synthetic topological insulator with tetO for chromatin-engineering (STITCH). By coupling STITCH with tetR linked to the KRAB domain to induce heterochromatin and disable the insulation, we developed a drug-inducible system to control gene activation by enhancers. We inserted STITCH into five different positions of the remaining allele of the locus: \\"STITCH+30kb\\", \\"STITCH+440kb\\", \\"STITCH+1760kb\\" and \\"STITCH+1790kb\\" have the STITCH insertions away from the MYC promoter for the indicated distances to the telomeric side of the p arm of the chromosome. \\"STITCH-30kb\\", at the 30-kb upstream from the MYC. We also made a deletion clone of the enhancer region, termed del(30-440). We made deletion of each CTCF array, L (delL) and R (delR), inversion of R (invR), deletion of the middle five binding sites from L2 to R2 (del(L2-R2)), and deletion of the six sites but for R3 (del(L1-R2)) in STITCH+30kb. We also obtained deletion and inversion of the whole of STITCH (del(L1-R3) and inv(L1-R3)). We integrated a transgene consisting of tetR-KRAB followed by DNA encoding the 2A peptide and the puromycin resistant gene with piggyBac transposition into the genome in the STITCH+30kb clone (STITCH/KRAB). We performed 4C-seq (Circular chromatin conformation capture assay followed by deep-sequencing) from the MYC promoter as a viewpoint to see how STITCH impacts on the chromatin conformation.