Journal: International Journal of Molecular Sciences

Article Title: Effective Restoration of Gastric and Esophageal Tissues in an In Vitro Model of GERD: Mucoadhesive and Protective Properties of Xyloglucan, Pea Proteins, and Polyacrylic Acid

doi: 10.3390/ijms26094409

Figure Lengend Snippet: Evaluation of barrier integrity of GTL-16 cells. ( A ) TEER (transepithelial electrical resistance) value measured using EVOM3 at different timepoints (from 1 h to 4 h); ( B – D ) analysis of TJ measured by Enzyme-Linked Immunosorbent Assay (ELISA) test (Claudin-1, ZO-1, and occludin, respectively) at 4 h. HCl = positive control, cells in GERD condition induced by HCl; Control = negative control, untreated cells. Data are expressed as means ± SD (%) of 5 independent experiments. * p < 0.05 vs. negative control; φ p < 0.05 vs. positive control. HCl: hydrochloric acid; TEER: trans-epithelial electrical resistance; XXPA: combination of xyloglucan, pea protein, and polyacrylic acid; ZO-1: Zonula Occludens-1.

Article Snippet: Fibronectin quantification was determined using the Human FN1 (Fibronectin) ELISA Kit (FineTest, Wuhan, China) according to the manufacturer’s instructions on GTL-16 cell lysates [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Positive Control, Control, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Effective Restoration of Gastric and Esophageal Tissues in an In Vitro Model of GERD: Mucoadhesive and Protective Properties of Xyloglucan, Pea Proteins, and Polyacrylic Acid

doi: 10.3390/ijms26094409

Figure Lengend Snippet: Adhesion study on GTL-16 cells. ( A ) Vitronectin and ( B ) fibronectin analysis measured by Enzyme-Linked Immunosorbent Assay (ELISA) test at 4 h. HCl = positive control, cells in GERD condition induced by HCl; Control = negative control, untreated cells. Data are expressed as means ± SD (%) of 5 independent experiments. * p < 0.05 vs. negative control; φ p < 0.05 vs. positive control. HCl: hydrochloric acid; XXPA: combination of xyloglucan, pea protein, and polyacrylic acid.

Article Snippet: Fibronectin quantification was determined using the Human FN1 (Fibronectin) ELISA Kit (FineTest, Wuhan, China) according to the manufacturer’s instructions on GTL-16 cell lysates [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Positive Control, Control, Negative Control

Journal: Molecules

Article Title: Vincamine Ameliorates Epithelial-Mesenchymal Transition in Bleomycin-Induced Pulmonary Fibrosis in Rats; Targeting TGF-β/MAPK/Snai1 Pathway

doi: 10.3390/molecules28124665

Figure Lengend Snippet: Effect of vincamine on fibronectin ( A ), N-cadherin ( B ), and collagen ( C ) levels in lung tissues. Bars represent mean ± SD. Significant difference was analyzed by one-way ANOVA test followed by post hoc Dunnett test, where * p < 0.001, compared to sham group, and # p < 0.001, compared to BLM-induced group.

Article Snippet: In lung tissue homogenates, levels of fibronectin, N-cadherin, and collagen were also assessed, according to the manufacturer’s instructions, utilizing rat fibronectin ELISA kit (#MBS761397, MyBioSource, CA, USA), N-cadherin ELISA kit (#KOA0665, Rockland immunochemical Inc., Royersford, PA, USA), and collagen type I ELISA kit (#MBS262647, MyBioSource, CA, USA), respectively.

Techniques:

Journal: Journal of Assisted Reproduction and Genetics

Article Title: GRN, NOTCH3, FN1, and PINK1 expression in eutopic endometrium – potential biomarkers in the detection of endometriosis – a pilot study

doi: 10.1007/s10815-020-01905-4

Figure Lengend Snippet: Dataset of conducted experiments with some descriptive statistics showing the sample size, median, range , and standard deviation

Article Snippet: The following kits were used: Progranulin ELISA Kit (No. E-EL-H1578, Elabscience, USA), Human Notch Homolog 3 ELISA Kit (No. SEL147Hu, Cloud-Clone Corp., USA), Quantikine ELISA Human Fibronectin (No. DFBN10, R&D Systems, USA), Human PINK1 ELISA Kit (No. MBS9327222, MyBioSource, USA).

Techniques: Standard Deviation, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Assisted Reproduction and Genetics

Article Title: GRN, NOTCH3, FN1, and PINK1 expression in eutopic endometrium – potential biomarkers in the detection of endometriosis – a pilot study

doi: 10.1007/s10815-020-01905-4

Figure Lengend Snippet: Changes in protein expression levels. Results of enzyme-linked immunosorbent assay (ELISA) of human eutopic endometrial tissue. Relative protein expression levels of GRN and FN1 to the total protein amount of the endometrial tissue sample. Bar graphs showing protein amount in pg (GRN) or ng (FN1) per 1 mg of total protein amount. Data represents the mean ± SEM. Control (without EM, n = 10); ASRM I (minimal EM, n = 10); ASRM II (mild EM, n = 10); ASRM III (moderate EM, n = 10). * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The following kits were used: Progranulin ELISA Kit (No. E-EL-H1578, Elabscience, USA), Human Notch Homolog 3 ELISA Kit (No. SEL147Hu, Cloud-Clone Corp., USA), Quantikine ELISA Human Fibronectin (No. DFBN10, R&D Systems, USA), Human PINK1 ELISA Kit (No. MBS9327222, MyBioSource, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: International journal of molecular sciences

Article Title: Endothelial Dysfunction and Impaired Wound Healing Following Radiation Combined Skin Wound Injury.

doi: 10.3390/ijms252312498

Figure Lengend Snippet: Figure 5. Effect of RCI on skin tissue endothelial dysfunction markers. Mice were subjected to sham injury, wounds alone, and 9.0 Gy RCI, and skin tissues surrounding the wounds were collected on days 3, 7, and 14 post-injury. Various markers associated with endothelial dysfunction were assessed in skin tissue lysates using ELISA. The markers analyzed include (Panel A) vascular endothelial growth factor (VEGF); (Panel B) Angiopoietin-2 (Ang2); (Panel C) intercellular adhesion molecule 1 (ICAM1); and (Panel D) insulin-like growth factor 1 (IGF1). Data are expressed as mean ± SEM. Significant differences in these markers were observed: * p < 0.05 for Sham vs. Wound/RCI on days 3, 7, and 14 post-injury, determined by one-way ANOVA followed by Dunnett’s multiple comparison test, and # p < 0.05 for Wound vs. RCI at the same time point, determined by two-way ANOVA followed by Sidak’s multiple comparison test. The sample size ranged from N = 3 to 9 per group per time point.

Article Snippet: The kits used were as follows: Mouse VEGF Immunoassay (MMV00-1, R&D Systems, Inc., Minneapolis, MN, USA), Mouse Angiopoietin-1 ELISA Kit (Colorimetric, NBP3-18634, Novus Biologicals, Centennial, CO, USA), Mouse/Rat Angiopoietin-2 Immunoassay (MANG20, R&D Systems, Inc., Minneapolis, MN, USA), Mouse/Rat IGF-I/IGF-1 Immunoassay (MG100, R&D Systems, Inc., Minneapolis, MN, USA), ICAM1 (CD54) Mouse ELISA Kit (ab100688, abcam, Waltham, MA, USA), Mouse TNF-α Immunoassay (MTA00B-1, R&D Systems, Inc., Minneapolis, MN, USA), Mouse CXCL1/KC Immunoassay (MKC00B-1, R&D Systems, Inc., Minneapolis, MN, USA), Bio-Plex ProTM TGF-β 3-plex assay (171W4001M, Bio-Rad Laboratories, Inc., Hercules, CA, USA), Mouse CTGF ELISA Kit (ab289838, abcam, Waltham, CA, USA), and Mouse Fibronectin ELISA Kit (NBP2-60517, Novus Biologicals, Centennial, CO, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: International journal of molecular sciences

Article Title: Endothelial Dysfunction and Impaired Wound Healing Following Radiation Combined Skin Wound Injury.

doi: 10.3390/ijms252312498

Figure Lengend Snippet: Figure 7. Effect of RCI on skin tissue levels of connective tissue growth factor (CTGF) and fibronectin. Mice were subjected to sham injury, wounds alone, and 9.0 Gy RCI, and skin tissues surrounding the wounds were collected on days 3, 7, and 14 post-injury. Levels of (Panel A) CTGF and (Panel B) fi- bronectin were measured in tissue lysates by EKISA. Data are expressed as mean ± SEM. Significant differences in these factors were observed: * p < 0.05 for Sham vs. Wound/RCI on days 3, 7, and 14 post-injury, determined by one-way ANOVA followed by Dunnett’s multiple comparison test, and # p < 0.05 for Wound vs. RCI at the same time point, determined by two-way ANOVA followed by Sidak’s multiple comparison test. The sample size ranged from N = 4 to 9 per group per time point.

Article Snippet: The kits used were as follows: Mouse VEGF Immunoassay (MMV00-1, R&D Systems, Inc., Minneapolis, MN, USA), Mouse Angiopoietin-1 ELISA Kit (Colorimetric, NBP3-18634, Novus Biologicals, Centennial, CO, USA), Mouse/Rat Angiopoietin-2 Immunoassay (MANG20, R&D Systems, Inc., Minneapolis, MN, USA), Mouse/Rat IGF-I/IGF-1 Immunoassay (MG100, R&D Systems, Inc., Minneapolis, MN, USA), ICAM1 (CD54) Mouse ELISA Kit (ab100688, abcam, Waltham, MA, USA), Mouse TNF-α Immunoassay (MTA00B-1, R&D Systems, Inc., Minneapolis, MN, USA), Mouse CXCL1/KC Immunoassay (MKC00B-1, R&D Systems, Inc., Minneapolis, MN, USA), Bio-Plex ProTM TGF-β 3-plex assay (171W4001M, Bio-Rad Laboratories, Inc., Hercules, CA, USA), Mouse CTGF ELISA Kit (ab289838, abcam, Waltham, CA, USA), and Mouse Fibronectin ELISA Kit (NBP2-60517, Novus Biologicals, Centennial, CO, USA).

Techniques: Comparison

Journal: Translational Vision Science & Technology

Article Title: Tissue Architecture Modulates Compositional and Structural Properties of Corneal Myofibroblast-Derived Matrix

doi: 10.1167/tvst.14.9.9

Figure Lengend Snippet: ( A ) Fibronectin deposition ( green ) and CHP ( magenta ) in aligned and random constructs at day14 in Z-stack images from whole mounts obtained with a confocal microscope. Scale bars : 20 µm. ( B – D ) ELISA assays were used to evaluate the protein concentration in relation to total protein content of the following proteins: fibronectin ( B ), decorin ( C ), and tenascin C ( D ). N = 32, derived from four donors, and each N represents the content from two construct lysates pooled together. Data presented as mean ± SD. ** P < 0.01; *** P < 0.001; **** P < 0.001; t P < 0.05; ### P < 0.001; #### P < 0.001; #, main effect of construct type.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits were used for detection of the following soluble protein fractions: collagen type I ( EKC33223 ; Biomatik, Kitchener, ON, Canada), collagen type V ( EKU11733 ; Biomatik), fibronectin (DFBN10; R&D Systems, Minneapolis, MN), tenascin C (EH446RB; Thermo Fisher Scientific), and decorin (Thermo Fisher Scientific) according to each manufacturer's instructions.

Techniques: Construct, Microscopy, Enzyme-linked Immunosorbent Assay, Protein Concentration, Derivative Assay

Journal: Breast Cancer Research : BCR

Article Title: A positive feedback loop driven by fibronectin and IL-1β sustains the inflammatory microenvironment in breast cancer

doi: 10.1186/s13058-023-01629-0

Figure Lengend Snippet: Expression of total fibronectin (tFN) and the alternatively spliced variant of cellular fibronectin harboring the extra domain A (FN-EDA) in breast cancer. A Gene expression of tFN and FN-EDA was evaluated in freshly obtained treatment-naïve breast tumor samples ( n = 62) by quantitative RT-PCR. Quantitation of the target gene expression was first normalized to the housekeeping β-actin gene expression for each sample. Then, the Ct values from the tumor-adjacent histopathologically normal breast tissue were used for additional normalization of the data. Results were distributed according to hormone receptor (HR) and HER2 status. B Kaplan–Meier plot for the estimation of survival in breast cancer patients ( n = 995, from METABRIC database) with low and high FN-EDA expression. C Immunofluorescence analysis of FN-EDA in the breast tumor specimens. Representative images of epithelial cellular adhesion molecule (EpCAM) and FN-EDA are demonstrated for each breast cancer subtype according to HR and HER2 expression. Large magnifications of the merged images are given as indents. The micrographs from one patient analyzed out of eight are shown for each subtype. (Scale bar, 10 µm). D Total FN and FN-EDA protein levels were determined in breast cancer cell lines by Western blot. E Change in tFN mRNA expression upon treatment with recombinant IL-1β, IL-6, or TNF-α in breast cancer cell lines was analyzed by RT-PCR. Quantitation of the target gene expression was first normalized to the housekeeping β-actin gene expression. Then, the Ct values from the control (untreated) cells were used for additional normalization. The dashed line crossing at 2. −ΔΔCt = 1 represents an equal expression level with the untreated control cells. The change in gene expression was calculated with 2e-ΔΔCt or -ΔΔCt formulas, which was previously described . The data from high-level fibronectin-expressing TNBC cancer cell lines MDA-MB-231 and HCC38 are shown in red. F The total FN secretion was assessed by ELISA and G the change in cell-associated (both intracellular and surface-bound) tFN levels were assessed by flow cytometry in the breast cancer cell lines stimulated with IL-1β, IL-6, or TNF-α for 24 h. H The impact of IL-1β on FN-EDA expression and cell morphology was assessed by immunofluorescence (scale bar, 40 µm). Representative micrographs are shown for MCF-7 and MDA-MB-231 cells. I Total FN and FN-EDA levels secreted into the culture media of breast cancer cell lines upon IL-1β treatment. Western blot bands were quantified, and the proportion of tFN to FN-EDA was shown in the bar graph on the right. J A correlation analysis between IL-1β and FN-EDA gene expression levels from breast tumor samples. The cell line data are from at least three independent experiments (Mean ± SEM, Student’s t test; *, P < 0.05; **, P < 0.01)

Article Snippet: ELISA kits for human fibronectin (Abcam) and IL-1β (BioLegend) were used according to the manufacturer’s instructions.

Techniques: Expressing, Variant Assay, Gene Expression, Quantitative RT-PCR, Quantitation Assay, Targeted Gene Expression, Immunofluorescence, Western Blot, Recombinant, Reverse Transcription Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Breast Cancer Research : BCR

Article Title: A positive feedback loop driven by fibronectin and IL-1β sustains the inflammatory microenvironment in breast cancer

doi: 10.1186/s13058-023-01629-0

Figure Lengend Snippet: The impact of FN-EDA on macrophage functions in breast cancer. A FN-EDA and CD68 immunofluorescence staining in HR- and HER2-positive breast cancer and in triple-negative breast cancer (TNBC) tissue (scale bar, 20 µm). B – D Freshly isolated monocytes were incubated for 48 h with CM from HR + breast cancer cell lines and from TNBC cell lines. The data from high-level fibronectin-expressing TNBC cancer cell lines MDA-MB-231 and HCC38 are shown in red. Expression of CD163, CD206, CD80, and CD86 macrophage differentiation and activation markers was assessed by flow cytometry. B Representative flow cytometry dot plots show the gating strategy for the monocytes treated with CM from HR + MCF-7 and TNBC MDA-MB-231 cells. C Percentage of CD163 + CD206 + macrophages and D median fluorescence intensity (MFI) value of CD80 and CD86 on CD163 + CD206 + cells. E Increasing amounts of monocytes treated with MCF-7 or MDA-MB-231 CM were co-cultured with T cells. T cell proliferation was assessed by flow cytometric CFSE dilution assay. F Phagocytosis, G production of reactive oxygen species, and H migration capacity of monocytes following the treatment with MCF-7 or MDA-MB-231 CM. I Polarization of monocytes on the surfaces coated with the plasma fibronectin (pFN), the recombinant FN lacking the EDA domain (rFN-EDA − ) and the recombinant FN with the EDA domain (rFN-EDA + ). The cells were pre-treated with CM from the TNBC cell lines. MDA-MB-468 cell line represents a TNBC cell line with low-level fibronectin expression (A.U, arbitrary units). Fluorescence microscopy images of actin staining for monocyte polarization analysis on different surfaces are presented on the right-hand side (scale bar, 40 µm). Each experiment was repeated at least three times with the monocytes from different donors. (Mean ± SEM, Student’s t test; *, P < 0.05; **, P < 0.01)

Article Snippet: ELISA kits for human fibronectin (Abcam) and IL-1β (BioLegend) were used according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Staining, Isolation, Incubation, Expressing, Activation Assay, Flow Cytometry, Fluorescence, Cell Culture, Dilution Assay, Migration, Clinical Proteomics, Recombinant, Microscopy

Journal: Breast Cancer Research : BCR

Article Title: A positive feedback loop driven by fibronectin and IL-1β sustains the inflammatory microenvironment in breast cancer

doi: 10.1186/s13058-023-01629-0

Figure Lengend Snippet: Association of FN-EDA with IL-1β and STAT3 inflammatory pathways. A Correlation of macrophage M2 subtype with FN-EDA expression in breast tumors assessed by using METABRIC data. B Phospho-STAT3 expression was determined in CD68 + macrophages in TNBC tumor specimens. Representative immunofluorescence staining from a patient is shown. C STAT3 phosphorylation (pSTAT3) levels were assessed by Western blot in the monocytes treated with CM from TNBC cell lines. MDA-MB-468 cell line represents the Basal-like A subtype of TNBC. D Correlation analysis of IL-1β and STAT3 gene expression in freshly obtained breast cancer tissues. E IL-1β secretion levels (upper panel) and NF-κB activity (lower panel) in the monocytes incubated with CM from TNBC cell lines in the presence of plasma fibronectin (pFN), recombinant FN lacking EDA domain (rFN-EDA − ) and recombinant FN with EDA domain (rFN-EDA + ) (A.U, arbitrary units). F Inhibition of TNBC CM-induced activation of the STAT3 pathway through stattic treatment was studied by Western blot. The control cells were treated with DMSO, the solvent for stattic. G Decrease in IL-1β secretion (upper panel) and NF-κB activity (lower panel) in the monocytes pre-treated with stattic. The fold change was calculated in comparison to the control monocytes incubated with CM from TNBC cell lines in the presence of the plasma fibronectin (pFN), the recombinant FN lacking the EDA domain (rFN-EDA − ) and the recombinant FN with the EDA domain (rFN-EDA. + ). (A.U, arbitrary units). Each experiment was repeated at least three times with monocytes from different donors. (Mean ± SEM, Student’s t test; *, P < 0.05; **, P < 0.01). (Black bars, monocytes in the control media; blue bars, CM from low-level fibronectin-expressing cell lines; red bars, CM from high-level fibronectin-expressing cell lines)

Article Snippet: ELISA kits for human fibronectin (Abcam) and IL-1β (BioLegend) were used according to the manufacturer’s instructions.

Techniques: Expressing, Immunofluorescence, Staining, Phospho-proteomics, Western Blot, Gene Expression, Activity Assay, Incubation, Clinical Proteomics, Recombinant, Inhibition, Activation Assay, Control, Solvent, Comparison

Journal: Cancers

Article Title: ROCK2 Confers Acquired Gemcitabine Resistance in Pancreatic Cancer Cells by Upregulating Transcription Factor ZEB1

doi: 10.3390/cancers11121881

Figure Lengend Snippet: Primer for PCR assay.

Article Snippet: Fibronectin was purchased from Proteintech Group Inc. P-ROCK1 (Thr455 + Ser456) was purchased from Bioss (Beijing, China).

Techniques: