Gene expression by NSC or SC osteoblasts.

<p>(A) less secreted proteins (B) more secreted proteins. The mRNA copy numbers were normalized against the corresponding copy number of HPRT mRNA and results are showed as the SC relative to NSC gene expression (mean ± SEM of five independent experiments performed with bone biopsy coming from different donors). In each experiment, each experimental condition was performed in duplicate. P: comparisons of mean values between NSC and SC were performed by unpaired t-test. Dotted blue line in each gene represents the result obtained with MS/MS in secretome with the corresponding protein.</p

Western-blotting detection of osteomodulin using goat polyclonal antisera raised against entire osteomodulin.

<p>(A)(B) rhOMD: human recombinant osteomodulin (positive control), DMEM: concentration unconditioned culture supernatant (negative control), NSC: non sclerotic osteoblasts supernatant (N36, N38, N40), SC: sclerotic osteoblasts supernatant (S36, S38, S40), C1-6: six healthy patients albumin/IgG depleted serum, OA1-6: six OA patients before total knee replacement surgery albumin/IgG depleted serum. (C) Quantification of osteomodulin western blot performed on osteoblasts culture supernatants and on albumin/IgG depleted serum of 6 healthy patients (controls) and 6 patients before total knee replacement (OA). Quantification was made using ImageJ on total band and normalized with total protein charged visualized with reverse total protein stain (Pierce).</p

OMD1 and OMD2 levels found in healthy and OA human sera.

<p>(A) OMD1, (B) OMD2. Data are presented as scatter dot plots with median from 22 patients in each group. Comparisons of values were performed by Mann Whitney test. (C) Spearman correlation between OMD1 and OMD2 in serum, n = 44.</p

Estimation of abundance of NSC osteoblasts secreted proteins using high top 3.

<p>Estimation of abundance of NSC osteoblasts secreted proteins using high top 3.</p

MS/MS proteomic analysis of osteoblasts secretome.

<p>MS/MS proteomic analysis of osteoblasts secretome.</p

OMD1 and OMD2 levels found in osteoblasts cell culture supernatant.

<p>Data are means +/- SEM from 10 patients. Comparisons of mean values were performed by unpaired t-test.</p

Yolk ferritin expression in reproductive organs of <i>L. stagnalis</i> exposed to testosterone (T), cyproterone acetate (CPA), chlordecone (CLD) and tributyltin (TBT) for 21 days.

<p>Results are expressed as percentage of expression measured in positive controls (egg samples) run with each gel. Error bars are SEM (<i>n</i>=4). Statistical differences from the controls; *: p<0.05. </p

Investigation of Structure-Stabilizing Elements in Proteins by Ion Mobility Mass Spectrometry and Collision-Induced Unfolding

A recently developed proteolytic reactor, designed for protein structural investigation, was coupled to ion mobility mass spectrometry to monitor collisional cross section (CCS) evolution of model proteins undergoing trypsin-mediated mono enzymatic digestion. As peptides are released during digestion, the CCS of the remaining protein structure may deviate from the classical 2/3 power of the CCS-mass relationship for spherical structures. The classical relationship between CCS and mass (CCS = A × M2/3) for spherical structures, assuming a globular shape in the gas phase, may deviate as stabilizing elements are lost during digestion. In addition, collision-induced unfolding (CIU) experiments on partially digested proteins provided insights into the CCS resilience in the gas phase to ion activation, potentially due to the presence of stabilizing elements. The study initially investigated a model peptide ModBea (3 kDa), assessing the impact of disulfide bridges on CCS resilience in both reduced and oxidized forms. Subsequently, β-lactoglobulin (2 disulfide bridges), calmodulin (Ca2+ coordination cation), and cytochrome c (heme) were selected to investigate the influence of common structuring elements on CCS resilience. CIU experiments probed the unfolding process, evaluating the effect of losing specific peptides on the energy landscapes of partially digested proteins. Comparisons of the TWCCSN2→He to trend curves describing the CCS/mass relationship revealed that proteins with structure-stabilizing elements consistently exhibit TWCCSN2→He and greater resilience toward CIU compared to proteins lacking these elements. The integration of online digestion, ion mobility, and CIU provides a valuable tool for identifying structuring elements in biopolymers in the gas phase

Baculoviral Protein and DNA Co-delivery System for Mammalian Genome Engineering - raw Sanger sequencing and flow-cytometry data

Raw Sanger sequencing (.ab1) and flow-cytometry (.FCS) data produced in the study Baculoviral Protein and DNA Co-delivery System for Mammalian Genome Engineering</p

Sparc-Like Protein 1 Is a New Marker of Human Glioma Progression

High-grade gliomas (glioblastomas) are the most common and deadly brain tumors in adults, currently with no satisfactory treatment available. Apart from <i>de novo</i> glioblastoma, it is currently accepted that these malignancies mainly progress from lower grade glial tumors. However, the molecular entities governing the progression of gliomas are poorly understood. Extracellular and membrane proteins are key biomolecules found at the cell-to-cell communication interface and hence are a promising proteome subpopulation that could help understand the development of glioma. Accordingly, the current study aims at identifying new protein markers of human glioma progression. For this purpose, we used glial tumors generated orthotopically with T98G and U373 human glioma cells in nude mice. This setup allowed also to discriminate the protein origin, namely, human (tumor) or mouse (host). Extracellular and membrane proteins were selectively purified using biotinylation followed by streptavidin affinity chromatography. Isolated proteins were digested and then identified and quantified employing 2D-nano-HPLC–MS/MS analysis. A total of 23 and 27 up-regulated extracellular and membrane proteins were identified in the T98G and U373 models, respectively. Approximately two-thirds of these were predominantly produced by the tumor, whereas the remaining proteins appeared to be mainly overexpressed by the host tissue. Following extensive validation, we have focused our attention on sparc-like protein 1. This protein was further investigated using immunohistochemistry in a large collection of human glioma samples of different grades. The results showed that sparc-like protein 1 expression correlates with glioma grade, suggesting the possible role for this protein in the progression of this malignancy