anti fibulin 5  (Danaher Inc)

Journal: bioRxiv

Article Title: The lung extracellular matrix protein landscape in severe early-onset and moderate chronic obstructive pulmonary disease

doi: 10.1101/2023.10.20.562391

Figure Lengend Snippet: FFPE lung tissue sections from control and COPD donors were stained using immunohistochemistry for ECM and ECM-associated proteins with specific signals being detected with Nova red (red) and counterstained with hematoxylin (blue). Sections were scanned at 40x magnification using a digital slide scanner. Scale bar = 400um. Images are representative of protein detection patterns seen in control donors (n=18), SEO-COPD (n=12), and moderate COPD (n=14). ECM: extracellular matrix; SEO-COPD: Severe early-onset COPD patients; FFPE: formalin fixed paraffin embedded; COL1A1: collagen type I α chain 1; COL6A1: collagen type VI α chain 1; COL6A2: collagen type VI α chain 2; COL14A1: collagen type XIV α chain 1; FBLN2: fibulin 2; FBLN5: fibulin 5; LTBP4: latent transforming growth factor binding protein 4; LUM: lumican; DCN: decorin; VCAN: versican; ELN: elastin.

Article Snippet: Following PBS washes, primary antibodies diluted in 1% BSA/PBS for FBLN5 (1:8000, Mouse Anti-Fibulin 5/DANCE Antibody 1G6A4, Novus Biologicals), DCN (1:1500, Mouse Anti-Dermatan Sulfate Proteoglycan Antibody 6B6, Seikagaku), VCAN (1:200, Mouse Anti-Versican Antibody 2B1, Seikagaku), and ELN (1:400, Rabbit Anti-Elastin Antibody CL55011AP, Cedarlane Labs) were added to the respective sections for 1 hour at room temperature.

Techniques: Control, Staining, Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Binding Assay

Journal: bioRxiv

Article Title: The lung extracellular matrix protein landscape in severe early-onset and moderate chronic obstructive pulmonary disease

doi: 10.1101/2023.10.20.562391

Figure Lengend Snippet: ECM proteins in FFPE tissue sections from controls (n=18), SEO-COPD (n=12), and moderate COPD (n=14) were detected using immunohistochemistry. Parenchyma was isolated and analyzed for percentage positive tissue area and mean intensity of positive pixels of expression for each protein. All 44 donors were available for analysis of COL1A1, COL6A2, FBLN2, LTBP4, LUM, DCN, VCAN, AND ELN, while for COL6A1, COL14A1, and FBLN5 43 donors were available. For each protein, regression coefficients (± 95% CI) were obtained following linear regression and a p value of <0.05 was considered significant. Non-transformed variables are plotted first, followed by log transformed variables. The differences in ECM and ECM-associated proteins in COPD tissue were compared to control in terms of A-C) percentage area and D-F) mean intensity of positively stained pixels. Dark blue-colored bars in light blue colored boxes highlight significant differences. ECM: extracellular matrix; SEO-COPD: Severe early-onset COPD patients; FFPE: formalin fixed paraffin embedded; COL1A1: collagen type I α chain 1; COL6A1: collagen type VI α chain 1; COL6A2: collagen type VI α chain 2; COL14A1: collagen type XIV α chain 1; FBLN2: fibulin 2; FBLN5: fibulin 5; LTBP4: latent transforming growth factor binding protein 4; LUM: lumican; DCN: decorin; VCAN: versican; ELN: elastin.

Article Snippet: Following PBS washes, primary antibodies diluted in 1% BSA/PBS for FBLN5 (1:8000, Mouse Anti-Fibulin 5/DANCE Antibody 1G6A4, Novus Biologicals), DCN (1:1500, Mouse Anti-Dermatan Sulfate Proteoglycan Antibody 6B6, Seikagaku), VCAN (1:200, Mouse Anti-Versican Antibody 2B1, Seikagaku), and ELN (1:400, Rabbit Anti-Elastin Antibody CL55011AP, Cedarlane Labs) were added to the respective sections for 1 hour at room temperature.

Techniques: Immunohistochemistry, Isolation, Expressing, Transformation Assay, Control, Staining, Formalin-fixed Paraffin-Embedded, Binding Assay

Journal: bioRxiv

Article Title: The lung extracellular matrix protein landscape in severe early-onset and moderate chronic obstructive pulmonary disease

doi: 10.1101/2023.10.20.562391

Figure Lengend Snippet: ECM proteins present in FFPE tissue sections from controls (n=18), SEO-COPD (n=12), and moderate COPD (n=14) were detected using immunohistochemistry. Airway walls were isolated and analyzed for percentage area and mean intensity of expression for each protein. The number of airway walls available for the analysis for each protein were COL1A1 (n= 155), COL6A1 (n=150), COL6A2 (n=149), COL14A1 (n=152), FBLN2 (n=158), FBLN5 (n=173), LTBP4 (n=163), LUM (n=158), DCN (n=156), VCAN (n=165), and ELN (n=158). For each protein, regression coefficients (± 95% CI) were obtained following linear regression and a p value of <0.05 was considered significant. Non-transformed variables are plotted first, followed by log transformed variables. The differences in ECM and ECM-associated proteins in COPD tissue were compared to control in terms of A-C) percentage area and D-F) mean intensity of positively stained pixels. Dark blue-colored bars in light blue colored boxes highlight significant differences. ECM: extracellular matrix; SEO-COPD: Severe early-onset COPD patients; FFPE: formalin fixed paraffin embedded; COL1A1: collagen type I α chain 1; COL6A1: collagen type VI α chain 1; COL6A2: collagen type VI α chain 2; COL14A1: collagen type XIV α chain 1; FBLN2: fibulin 2; FBLN5: fibulin 5; LTBP4: latent transforming growth factor binding protein 4; LUM: lumican; DCN: decorin; VCAN: versican; ELN: elastin.

Article Snippet: Following PBS washes, primary antibodies diluted in 1% BSA/PBS for FBLN5 (1:8000, Mouse Anti-Fibulin 5/DANCE Antibody 1G6A4, Novus Biologicals), DCN (1:1500, Mouse Anti-Dermatan Sulfate Proteoglycan Antibody 6B6, Seikagaku), VCAN (1:200, Mouse Anti-Versican Antibody 2B1, Seikagaku), and ELN (1:400, Rabbit Anti-Elastin Antibody CL55011AP, Cedarlane Labs) were added to the respective sections for 1 hour at room temperature.

Techniques: Immunohistochemistry, Isolation, Expressing, Transformation Assay, Control, Staining, Formalin-fixed Paraffin-Embedded, Binding Assay

Journal: bioRxiv

Article Title: The lung extracellular matrix protein landscape in severe early-onset and moderate chronic obstructive pulmonary disease

doi: 10.1101/2023.10.20.562391

Figure Lengend Snippet: ECM differences noted in the different analyzes throughout this study have been summarized here. In the staining and ECM signatures, red or blue arrows indicate higher or lower proportional levels or component scores in COPD respectively, while they denote positive or negative associations with FEV1 respectively. ECM: extracellular matrix; SEO-COPD: Severe early-onset COPD patients; COL1A1: type I collagen α chain 1; COL6A1: type VI collagen α chain 1; COL6A2: type VI collagen α chain 2; COL14A1: type XIV collagen α chain 1; FBLN2: fibulin 2; FBLN5: fibulin 5; LTBP4: latent transforming growth factor binding protein 4; LUM: lumican; DCN: decorin; VCAN: versican; ELN: elastin; FEV1: forced expiratory volume in 1 second.

Article Snippet: Following PBS washes, primary antibodies diluted in 1% BSA/PBS for FBLN5 (1:8000, Mouse Anti-Fibulin 5/DANCE Antibody 1G6A4, Novus Biologicals), DCN (1:1500, Mouse Anti-Dermatan Sulfate Proteoglycan Antibody 6B6, Seikagaku), VCAN (1:200, Mouse Anti-Versican Antibody 2B1, Seikagaku), and ELN (1:400, Rabbit Anti-Elastin Antibody CL55011AP, Cedarlane Labs) were added to the respective sections for 1 hour at room temperature.

Techniques: Staining, Binding Assay

Journal: Food Science & Nutrition

Article Title: Anti‐skin aging effect of sea buckthorn proanthocyanidins in D‐galactose‐induced aging mice

doi: 10.1002/fsn3.3823

Figure Lengend Snippet: The effect of SBP on Col I and tropoelastin synthesis pathways. (a) Associated protein bands. (b, c) Expression of TGF‐β1, Smad3, Col I, Elastin, LOX, and Fibulin 5 in skin tissues of different experimental groups. Protein blots are representative of at least three independent experiments. (d) Scanning electron microscopic observation of fibrous tissue in the dermis of mice skin tissue. a‐f Mean values within the same row not sharing a common superscript letter are significantly different ( p < .05) by Duncan's multiple range test.

Article Snippet: Primary antibodies: anti‐Smad 3 was purchased from Proma (Hunan, China, cat. no. 30072); anti‐TGF‐β1 was purchased from ABclonal (Wuhan, China, cat. no. A2124); and anti‐type I collagen (cat. no. ab260043), anti‐tropoelastin (cat. no. Ab217356), anti‐LOX (cat. no. ab174316), and anti‐Fibulin 5 (cat. no. ab109428) were purchased from Abcam (Cambridge, MA, USA). β‐Actin was purchased from Protein Group, Inc. (Wuhan, China, cat. no. 66009‐1‐Ig).

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Proteomic mapping reveals dysregulated angiogenesis in the cerebral arteries of rats with early-onset hypertension

doi: 10.1016/j.jbc.2023.105221

Figure Lengend Snippet: List of significantly regulated proteins detected in cerebral arteries from hypertensive rats compared to normotensive control

Article Snippet: Tissue sections were blocked in blocking buffer (5% normal swine serum (Jackson ImmunoResearch), 1% bovine serum albumin (BSA, Sigma), 0.1% TritonX-100 (Sigma) in 1 × PBS) and stained with commercial anti-alpha smooth muscle actin antibody [1A4] (ab7817, 1:400, Abcam), anti-Fibulin 5 antibody (ab202977, 1:200, Abcam), and anti-human cadherin-13 antibody (AF3264, 1:200, R&D Systems) diluted in 1% BSA, 0.1% TritonX-100 in 1×PBS overnight at 4 °C.

Techniques: Molecular Weight, Membrane

Journal: The Journal of Biological Chemistry

Article Title: Proteomic mapping reveals dysregulated angiogenesis in the cerebral arteries of rats with early-onset hypertension

doi: 10.1016/j.jbc.2023.105221

Figure Lengend Snippet: Differential expression of two candidate proteins across vascular beds. A , Venn diagram showing number of angiogenesis-associated proteins in cerebral arteries that are shared with mesenteric and renal arteries but change in opposite direction. B , column graph depicting logFC difference of Fbln5 (= blue ) and Cdh13 (= gray ) in cerebral, renal, and mesenteric artery comparisons between 12-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). C – G , immunohistochemistry analysis of α-SMA ( green ), Fbln5 ( red ), Cdh13 ( magenta ) in cerebral arteries from WKY controls and SHRs (n = 4 biological replicates, 2–4 technical replicates per rat). Scale bar represents 20 μm. Max fluorescent intensity was measured across a line. SEM is included on column graphs (WKY = blue , SHR = orange ).

Article Snippet: Tissue sections were blocked in blocking buffer (5% normal swine serum (Jackson ImmunoResearch), 1% bovine serum albumin (BSA, Sigma), 0.1% TritonX-100 (Sigma) in 1 × PBS) and stained with commercial anti-alpha smooth muscle actin antibody [1A4] (ab7817, 1:400, Abcam), anti-Fibulin 5 antibody (ab202977, 1:200, Abcam), and anti-human cadherin-13 antibody (AF3264, 1:200, R&D Systems) diluted in 1% BSA, 0.1% TritonX-100 in 1×PBS overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry

Journal: Biomedicines

Article Title: Ten-Eleven Translocation 1 and 2 Enzymes Affect Human Skin Fibroblasts in an Age-Related Manner

doi: 10.3390/biomedicines11061659

Figure Lengend Snippet: Effects of TET1 and TET2 on Fibulin 5 in fibroblasts obtained from young ( n = 5) and age-advanced ( n = 5) study participants. ( A ) TET1 overexpression-related changes in the expression of Fibulin 5 and percentage of cells expressing it. ( B ) TET2 overexpression-related changes in the expression of Fibulin 5 and percentage of cells expressing it. Statistical analysis was performed using the Mann-Whitney U test (young pSELECT-GFPzeo vs. age-advanced pSELECT-GFPzeo) and Wilcoxon matched-pairs signed rank test (pSELECT-GFPzeo vs. pSELECT-GFPzeo-TET1 or pSELECT-GFPzeo-TET2). MFI—mean fluorescence intensity.

Article Snippet: Forty-eight hours after transfection, cells were fixed with Cytofix/Cytoperm™ (BD Biosciences) and incubated with anti-Beclin 1 (1:200; Abcam, Cambridge, UK) or anti-Fibulin 5 (1:200; Biorbyt, Cambridge, UK) antibodies for 40 min. Next, the cells were incubated with secondary antibodies, as appropriate, for 30 min.

Techniques: Over Expression, Expressing, MANN-WHITNEY, Fluorescence

Journal: Biomedicines

Article Title: Ten-Eleven Translocation 1 and 2 Enzymes Affect Human Skin Fibroblasts in an Age-Related Manner

doi: 10.3390/biomedicines11061659

Figure Lengend Snippet: Summary of the effects of TET1 and TET2 on fibroblasts isolated from the skin of young and age-advanced individuals.

Article Snippet: Forty-eight hours after transfection, cells were fixed with Cytofix/Cytoperm™ (BD Biosciences) and incubated with anti-Beclin 1 (1:200; Abcam, Cambridge, UK) or anti-Fibulin 5 (1:200; Biorbyt, Cambridge, UK) antibodies for 40 min. Next, the cells were incubated with secondary antibodies, as appropriate, for 30 min.

Techniques: Isolation, BrdU Incorporation Assay, Activity Assay