Journal:

Article Title: A mutant, noninhibitory plasminogen activator inhibitor type 1 decreases matrix accumulation in experimental glomerulonephritis

doi: 10.1172/JCI200318038

Figure Lengend Snippet: Immunofluorescent staining score for ECM proteins at d6. Glomerular staining for FN-EDA+ (a), laminin (b), type I collagen (c), and type III collagen (d) were lower in the PAI-1R–treated, nephritic group. *P < 0.001 compared with normal control. #P < 0.01 compared with disease control.

Article Snippet: Monoclonal mouse anti-cellular fibronectin extra domain positive (FN-EDA+) (Harlan Sera-Lab Ltd., Loughborough, United Kingdom), rabbit anti–mouse laminin (ICN Immunobiologicals, Aurora, Ohio, USA), goat anti–human type I collagen, and goat anti–human type III collagen (Southern Biotechnology Associates, Birmingham, Alabama, USA) were used as the primary Ab’s for detection of matrix components in glomeruli at d6.

Techniques: Staining

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Schematic representation of the human collagen XVIII variants, termed as SHORT, MIDDLE and LONG/FZ. Collagenous sequences are shown in white. Non-collagenous (NC) amino terminal sequences common to all variants are shown in black. Non-collagenous amino terminal sequences common to the two long variants are shown in grey. A non-collagenous amino terminal sequence specific to the LONG/FZ variant is marked with waves. The amino acid lengths (aa) and the predicted molecular masses (kDa) of these sequences are given, as are those of the full length type XVIII collagen molecules. Signal sequences are indicated by vertical and horizontal hatching. The epitopes of the antibodies used in this work are shown by black horizontal lines. The molecular masses of some endostatin-containing carboxy-terminal fragments are also indicated.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Sequencing, Variant Assay

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Immunoprecipitation with anti-ALL antibody and detection with anti-LONG huXVIII antibody. Elevated type XVIII collagen precursors in the plasma of ALI patients compared with normal controls is shown. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Immunoprecipitation, Clinical Proteomics

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Western blot by anti-ALL huXVIII antibody. It shows elevated type XVIII collagen precursors in the bronchoalveolar lavage fluid (BALF) of patients with acute lung injury compared with normal controls. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Western Blot

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 expression is increased in lung cancer and associated with poor prognosis. Expression levels of COL3A1 in LUAD and LUSC assessed using the UALCAN (a) and GEPIA (b) databases. (c) Associations between COL3A1 expression levels and OS of patients with lung cancer, LUAD, and LUSC using kmplot.

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: Expressing

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 promotes cell viability and inhibits cell apoptosis in lung cancer. The mRNA and protein levels of COL3A1 in BEAS-2B, A549, H1299, PC-9, and NCI-H1703 cells detected using qRT-PCR (a) and Western blotting (b) (## P < 0.01). (c) Transfected efficiencies of shCOL3A1 and COL3A1 detected by Western blotting in A549 and H1299 cells (## P < 0.01, ### P < 0.001, vs. NC group; ** P < 0.01, vs. shNC group). (d) COL3A1 effect on cell growth using CCK-8 assay (## P < 0.01 vs. NC group; ** P < 0.01 vs. shNC group). (e) COL3A1 effect on cell viability using colony formation assay (### P < 0.001 vs. NC group; ** P < 0.01 vs. shNC group). (f) Effects of COL3A1 on cell apoptosis on flow cytometry assay (## P v 0.01, ### P < 0.001 vs. NC group; ** P < 0.01, *** P < 0.001 vs. shNC group).

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: Quantitative RT-PCR, Western Blot, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 promotes DDP resistance in lung cancer. (a) IC50 of A549/DDP and H1299/DDP cells to DDP assessed using the CCK-8 assay. (b) Effects of COL3A1 downregulation on the IC50 of A549/DDP and H1299/DDP to DDP assessed by the CCK-8 assay. (c) Effects of COL3A1 downregulation on the clone formation ability of A549/DDP and H1299/DDP to DDP assessed by the clone formation assay (### P < 0.001). (D) Effects of COL3A1 downregulation on the apoptosis of A549/DDP and H1299/DDP to DDP assessed by the flow cytometry assay (### P < 0.001).

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: CCK-8 Assay, Tube Formation Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Small-Molecule-Mediated Suppression of BMP Signaling by Selective Inhibition of BMP1-Dependent Chordin Cleavage

doi: 10.3390/ijms24054313

Figure Lengend Snippet: NPL1010 and NPL3008 rarely inhibit BMP1-dependent COL3A1 cleavage. ( A ) COL3A1 cleavage assay. rhCOL3A1 was incubated with rhBMP1. The effects of NPL1010 and NPL3008 on BMP1-dependent rhCOL3A1 cleavage were analyzed using SDS–PAGE and Oriole staining. Arrows indicate full-length rhCOL3A1 and cleaved rhCOL3A1. ( B ) Statistical analysis of BMP1-dependent cleavage of COL3A1. The levels of noncleaved and cleaved COL3A1 were measured by performing densitometry of the Oriole-stained gel, and the percentage of cleaved COL3A1 was calculated based on the fragments detected at each molecular weight and background. **: p < 0.01 (Dunnett’s test). The error bar indicates +s.d. with individual data shown ( n = 4).

Article Snippet: Recombinant human BMP1 (rhBMP1, 0.276 mg/mL, dissolved in 25 mM HEPES containing 400 mM ammonium sulfate, R&D Systems, Minneapolis, MN, USA, catalog no. 1927-ZN-010) and recombinant human COL3A1 (rhCOL3A1, 0.336 mg/mL, dissolved in 25 mM sodium acetate containing 1 M NaCl, R&D Systems, catalog no. 7294-CL-020) were divided into aliquots and stored at −80 °C.

Techniques: Cleavage Assay, Incubation, SDS Page, Staining, Molecular Weight

Journal: Orphanet journal of rare diseases

Article Title: Dysregulation of extracellular matrix and Lysyl Oxidase in Ehlers-Danlos syndrome type IV skin fibroblasts.

doi: 10.1186/s13023-023-03007-7

Figure Lengend Snippet: Fig. 1 Collagen IIIA protein expression in wild type and vEDS fibroblast cell lysate. Primary human dermal fibrobasts derived from normal (WT) or vEDS individuals were grown in complete media for 2 weeks, washed with PBS and lysed in RIPA buffer and normalized to total protein content. Samples were analyzed for Collagen III protein concentration using commercially available ELISA kit. Average values are reported from triplicate measurements with error bars representing STDEV

Article Snippet: The samples were tested using a standard sandwich enzyme-linked immunosorbent assay for the quantitative detection of human Collagen III according to manufactures instructions. (Aviva Systems, Cat#OKEH00269).

Techniques: Expressing, Derivative Assay, Protein Concentration, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Regenerative Skin Remodeling by a Dual Hyaluronic Acid Hybrid Complex in Multimodal Preclinical Models

doi: 10.3390/ijms27021027

Figure Lengend Snippet: Collagen and elastin production in HDFs treated with DHC and other comparators. ( A ) Collagen I, ( B ) Collagen III, and ( C ) Elastin levels were quantified by ELISA. Values are expressed as mean ± SD. * p < 0.05 vs. control; # p < 0.05 between groups.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) was designed to quantify the production of wrinkle-improving proteins using: Human Pro-Collagen I α1 SimpleStep ELISA Kit (ab210966, Abcam, Cambridge, MA, USA), Human Collagen Type III Alpha 1 ELISA Kit (NBP2-75858, Novus Biologicals, Centennial, CO, USA), Human Elastin ELISA Kit (MBS704171, MyBioSource, San Diego, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 expression is increased in lung cancer and associated with poor prognosis. Expression levels of COL3A1 in LUAD and LUSC assessed using the UALCAN (a) and GEPIA (b) databases. (c) Associations between COL3A1 expression levels and OS of patients with lung cancer, LUAD, and LUSC using kmplot.

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: Expressing

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 promotes cell viability and inhibits cell apoptosis in lung cancer. The mRNA and protein levels of COL3A1 in BEAS-2B, A549, H1299, PC-9, and NCI-H1703 cells detected using qRT-PCR (a) and Western blotting (b) (## P < 0.01). (c) Transfected efficiencies of shCOL3A1 and COL3A1 detected by Western blotting in A549 and H1299 cells (## P < 0.01, ### P < 0.001, vs. NC group; ** P < 0.01, vs. shNC group). (d) COL3A1 effect on cell growth using CCK-8 assay (## P < 0.01 vs. NC group; ** P < 0.01 vs. shNC group). (e) COL3A1 effect on cell viability using colony formation assay (### P < 0.001 vs. NC group; ** P < 0.01 vs. shNC group). (f) Effects of COL3A1 on cell apoptosis on flow cytometry assay (## P v 0.01, ### P < 0.001 vs. NC group; ** P < 0.01, *** P < 0.001 vs. shNC group).

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: Quantitative RT-PCR, Western Blot, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Balkan Medical Journal

Article Title: COL3A1 Overexpression Associates with Poor Prognosis and Cisplatin Resistance in Lung Cancer

doi: 10.4274/balkanmedj.galenos.2022.2022-6-16

Figure Lengend Snippet: COL3A1 promotes DDP resistance in lung cancer. (a) IC50 of A549/DDP and H1299/DDP cells to DDP assessed using the CCK-8 assay. (b) Effects of COL3A1 downregulation on the IC50 of A549/DDP and H1299/DDP to DDP assessed by the CCK-8 assay. (c) Effects of COL3A1 downregulation on the clone formation ability of A549/DDP and H1299/DDP to DDP assessed by the clone formation assay (### P < 0.001). (D) Effects of COL3A1 downregulation on the apoptosis of A549/DDP and H1299/DDP to DDP assessed by the flow cytometry assay (### P < 0.001).

Article Snippet: The scramble control (NC and shNC), lentiviral vector COL3A1 (catalog id: RC211766L3V), and COL3A1 shRNAs (shCOL3A1, catalog id: TL313797V) were bought from Origene (Beijing, China).

Techniques: CCK-8 Assay, Tube Formation Assay, Flow Cytometry