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: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: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: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:This is a TMT6 sample containing the following sub-samples:
TAG 126: standard, used for normalization purposes, It is a mixture of different biological samples.
TAG 127: Lysate of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
TAG 128: Lysate of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium (mixed human and mouse cell population).
TAG 129: Lysate of naive human pluripotent stem cells cultured on VTN coating in RSeT medium.
TAG 130: Not relevant.
TAG 131: standard, used for normalization purposes, It is a mixture of different biological samples. Same as sub-sample with tag 126.
Detailed culture and processing methods are described in Cesare et al, under submission.
Project description:Quantitative proteomics results of RTNs-KD neurons or DP1-KD neurons (DIV5). Peptides derived from Untreated neurons (UNT) were chemically labeled with TMT-126, pSuper-neurons with TMT-127, shRNAs_RTNs-neurons with TMT-128 and shRNA_DP1-neurons with TMT-131.
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.