Journal: Arthritis Research & Therapy

Article Title: Anti-type II collagen immune complex-induced granulocyte reactivity is associated with joint erosions in RA patients with anti-collagen antibodies

doi: 10.1186/s13075-015-0523-7

Figure Lengend Snippet: Functional reactivity against IgG as part of surface-bound collagen type II autoantibody (anti-CII) immune complex and polyclonal IgG. Amounts and functional reactivity against IgG as part of surface-bound collagen type II autoantibody (anti-CII) immune complex and polyclonal IgG directly coated to identical enzyme-linked immunosorbent assay (ELISA) plates. (a) ELISA reactivity of IgG against type II collagen (CII) aligned to known concentrations of directly coated IgG; these anti-CII levels (μg/ml) were thereafter used in the comparisons below. (b) Peripheral blood mononuclear cell-derived tumor necrosis factor alpha (ΤΝFα). (c) , (d) Polymorphonuclear granulocyte expression of CD66b and of CD16 respectively were compared for different concentrations of anti-CII and polyclonal IgG. Panels depict one representative result out of two to four experiments performed for each comparison. MFI, mean fluorescence intensity; OD, optical density.

Article Snippet: ELISA plates were coated with native human CII (10 μg/ml) (ELISA grade; Chondrex Inc.) at 4°C overnight.

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Fluorescence

Journal: bioRxiv

Article Title: Single cell landscape of hypertrophic scars identifies serine proteases as key regulators of myofibroblast differentiation

doi: 10.1101/2020.06.17.157073

Figure Lengend Snippet: A,B) Western blot of primary FBs stimulated with active TGFβ1 for 24h to differentiate FBs into alpha-smooth muscle actin-expressing (αSMA) myofibroblasts. Myofibroblast differentiation inhibited with A) DPP4-inhibitor Sitagliptin or B) urokinase-inhibitor BC-11. C, D) Collagen I or E, F) fibronectin in supernatants of stimulated primary skin FBs, detected by Enzyme-linked Immunosorbent Assay (ELISA). Whiskers represent range maximum and minimum values with < 1.5 interquartile range, boxes represent 25 th −75 th quartiles, line represents mean. Statistical significance was tested using one-way ANOVA with Tukey post-test. NS p>0.05, *p<0.05, **p<0.01, ***p<0.001. Experiments were performed in duplicates of five donors each.

Article Snippet: Human pro-collagen Ia1 ELISA (R&D Systems) and human fibronectin ELISA (R&D Systems) were performed with supernatants of TGFβ 1-stimulated FBs according to the manufacturer’s manual.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Characterization of bursa subacromialis-derived mesenchymal stem cells

doi: 10.1186/s13287-015-0104-3

Figure Lengend Snippet: Primer sequences and PCR conditions

Article Snippet: Immunohistochemical detection of collagen type II (COL II) was performed as described in detail previously [ ] using a primary monoclonal COL II antibody (Acris Antibodies GmbH, Herford, Germany), while negative controls were treated with mouse serum instead.

Techniques: Sequencing, Marker

Journal: Stem Cell Research & Therapy

Article Title: Characterization of bursa subacromialis-derived mesenchymal stem cells

doi: 10.1186/s13287-015-0104-3

Figure Lengend Snippet: Chondrogenic differentiation of BS cells and BMSCs. Cells cultivated in chondrogenic medium showed strong staining for proteoglycans determined by positive alcian blue staining (Alc Blue) and were also positive for collagen type II (COL II) in BS cells as well as in BMSC compared to cells cultivated in control medium ( a ). Left scale bar = 200 μm; right scale bar = 100 μm. b Expression of chondrogenic marker genes was evaluated using RT-PCR. Cultivation in the presence of chondrogenic medium (Ch) resulted in an increased expression of aggrecan (AGN), decorin (DEC), SRY (sex determining region Y)-box 9 (SOX9), indian hedgehog (IHH), and collagen type II (COL II) as compared to untreated cells (Co) for both cell types. Expression levels of the housekeeping gene elongation factor 1α (EF1α) are shown in the last row. Representative images from three different donors are shown. BMSCs bone-marrow derived mesenchymal stem cells BS bursa subacromialiss

Article Snippet: Immunohistochemical detection of collagen type II (COL II) was performed as described in detail previously [ ] using a primary monoclonal COL II antibody (Acris Antibodies GmbH, Herford, Germany), while negative controls were treated with mouse serum instead.

Techniques: Staining, Expressing, Marker, Reverse Transcription Polymerase Chain Reaction, Derivative Assay