Journal: Molecular Cancer

Article Title: c-Rel drives pancreatic cancer metastasis through fibronectin-integrin signaling-induced isolation stress resistance and EMT

doi: 10.1186/s12943-025-02486-5

Figure Lengend Snippet: A Representative images of Masson's trichrome (p<0.0001) and Picosirius-red stained tumors and their quantification (p=0.0248) B Representative FN1-IHC images (p=0.0186). GCKP-1 reflects fibrillar FN1 deposition around cancer cells individually. GCKP-2 reflects the perinuclear FN1 signal intracellularly, as shown with arrowheads. C Western blot for FN1 in bulk tumors and isolated cancer cell lysates. The expression is normalized to HSP90 (tissues p=0.0002, cells n.s.) D Mouse serum FN1 ELISA results (One-way ANOVA for a linear trend p=0.0366) E Fibronectin adhesion assay. The number of cells attached to the FN1-coated wells was normalized to the seeding number (One-way ANOVA test for a linear trend p=0.0242) F Heatmap of the differentially expressed genes in the RNA-seq analysis of CCKP-CKP-GCKP cells. G Principal component analysis (PCA) plot of the cells used for the RNAseq analysis. H Venn diagrams illustrating the number of common genes up- or down-regulated in the given comparison pairs. I GSEA of the DEGs between the given comparison pairs. The red stars indicate enrichments related to ECM remodeling, whereas the blue stars indicate EMT-related pathways. Unless otherwise indicated, for all analyses, ordinary one-way ANOVA was used. ANOVA p values are given in the figure legends. Tukey's multiple comparison tests are shown in the figures

Article Snippet: The serum samples were diluted 1:8000 and analyzed with a Mouse Fibronectin ELISA Kit (Novus, NBP2-60517).

Techniques: Staining, Western Blot, Isolation, Expressing, Enzyme-linked Immunosorbent Assay, Cell Adhesion Assay, RNA Sequencing, Comparison

Journal: Molecular Cancer

Article Title: c-Rel drives pancreatic cancer metastasis through fibronectin-integrin signaling-induced isolation stress resistance and EMT

doi: 10.1186/s12943-025-02486-5

Figure Lengend Snippet: A Colony-forming frequency was assessed by the ability of cells to form an organoid in Matrigel (one-way ANOVA, p=0.0015). Tukey's multiple comparisons test results are shown in the figure. B Representative microscopy images of spheroids formed under nonadherent conditions. The average spheroid diameter was quantified and analyzed (genotype factor p=0.0227). C Representative spheroid images and their diameter quantification. The spheroid medium was supplemented with either human plasma fibronectin (pFN1) or cellular fibronectin with EDB domain + type II domains 8−14 (EDB cFN1.3) (treatment factor p= n.s.) D Surface expression of selected proteins on spheroids as assessed by flow cytometry analysis (one-way ANOVA test for CD61 p=0.0244, CD133, EpCAM, CD44, CXCR4, Sca1 p= n.s.). Tukey's multiple comparisons test results are shown in the figure. E Representative macroscopic and microscopic images of the transplanted mouse lungs. The number of cell lines used per genotype are CKP=5, GCKP=5 and FNGCKP=4. Each cell line was injected two times to two individual mice. Nested one-way ANOVA was used for both analyses. Tukey's multiple comparisons test results are shown in the figure. Unless otherwise indicated, for all analyses, two-way ANOVA was used. ANOVA p values are given in the figure or their legends. Šídák's multiple comparison tests between genotypes are shown in the figures. F Schematic depicting the function of c-Rel in PDAC pathophysiology. c-Rel is an oncogenic factor involved in PDAC survival and metastasis. Increased c-Rel expression remodels the ECM by converting the collagen-rich stroma to a fibronectin-rich stroma. Cancer cells undergo epithelial-to-mesenchymal transition, increasing EMP by reducing the surface expression of E-cadherin and contractility via Src and Stat3 phosphorylation. Once isolation stress is induced under nonadherent conditions, c-Rel induces a niche enriched with fibronectin, coupled with increased Itgb3, Itga5 and Itgav expression. These changes are also supported by EMP, where c-Rel directly induces EMT-TFs such as Snai1, Snai2, Zeb2, and Tgfb . Increased tolerance to isolation stress ultimately drives metastasis. Created in BioRender. Kabacaoglu, D. (2025)

Article Snippet: The serum samples were diluted 1:8000 and analyzed with a Mouse Fibronectin ELISA Kit (Novus, NBP2-60517).

Techniques: Microscopy, Clinical Proteomics, Expressing, Flow Cytometry, Injection, Comparison, Phospho-proteomics, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Heart Failure Promotes Cancer Progression in an Integrin β1-Dependent Manner

doi: 10.3390/ijms242417367

Figure Lengend Snippet: TAC-operation promotes LLC cancer cell growth in periostin KO mice. ( A ) Serum levels of B6 (green) and POSTN (−/−) (red) mice ( n = 3 per group) were obtained by ELISA for Periostin. ( B ) Schematic experimental timeline for the Lewis lung carcinoma (LLC) cell model implanted in Periostin KO (POSTN (−/−) ) male mice. ( C ) POSTN (−/−) mice were subcutaneously implanted into the flanks with LLC cells (0.5 × 106 cells per mouse) ( n = 7). Mice were divided into 2 groups: TAC-operated group (red, n = 3), and non-opereted group (black, n = 4). Tumors were monitored over time and tumor volume was calculated using the formula: Width × Length × 0.5. ( D ) Tumor weight at sacrifice. Each dot represents one mouse. Data are presented as mean ± SE. Two-way repeated measures ANOVA followed by the Bonferroni post-test ( C ), and Student’s t -test ( A , D ). * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: The quantification of Periostin and Fibronectin in the serum was performed using the mouse Periostin/OSF-2 Quantikine ELISA kit (R&D systems Inc., Minneapolis, MN, USA) and mouse Fibronectin ELISA kit (E-EL-M0506, Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart Failure Promotes Cancer Progression in an Integrin β1-Dependent Manner

doi: 10.3390/ijms242417367

Figure Lengend Snippet: Tumor-promotion phenotype in the serum derived from POSTN (−/−) mice is mediated by multiple secreted factors. The proliferation of ( A ) polyoma middle T breast cancer cells (PyMT) or ( B ) Lewis lung carcinoma (LLC) cells under different growth conditions in the presence of the indicated murine serums; n = 4 plates per treatment for each time point. ( C ) Pooled serum from POSTN (−/−) or C57Bl/6 male mice ( n = 3 per pool) was used to probe the proteome cytokine array. For each protein, the serum levels are presented as a fold change in the serum derived from POSTN (−/−) relative to the control C57Bl/6 serum. ( D ) Fibronectin (FN) serum levels measured by ELISA ( n = 3 per group; C57Bl/6 in green, POSTN (−/−) in red). Data are presented as mean ± SE. One-way ANOVA followed by Tukey’s post-test ( A , B ), and Student’s t -test ( D ). * p < 0.05, ** p < 0.01.

Article Snippet: The quantification of Periostin and Fibronectin in the serum was performed using the mouse Periostin/OSF-2 Quantikine ELISA kit (R&D systems Inc., Minneapolis, MN, USA) and mouse Fibronectin ELISA kit (E-EL-M0506, Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Tenomodulin is essential for prevention of adipocyte accumulation and fibrovascular scar formation during early tendon healing

doi: 10.1038/cddis.2017.510

Figure Lengend Snippet: The absence of Tnmd increases erroneous ECM deposition. ( a , b ) Immunofluorescence staining with anti-Tnmd C-terminal antibody showed Tnmd secretion in the ECM of WT Achilles tendon, but not in Tnmd −/− . ( c–h ) Biglycan, Comp and fibronectin protein deposition in the tendon scar, analyzed by fluorescent digital signal quantification, was clearly augmented in Tnmd −/− when compared with WT mice at 8 days postoperatively. ( i–l ) Biglycan and fibronectin protein levels from cell lysates and supernatant were assessed by ELISA and the levels of both proteins were significantly increased in the supernatant and slightly increased in the cell lysates of Tnmd −/− versus WT TSPCs. For quantification in ( b , d , f and h ), statistical significance was calculated using two-tailed non-parametric Mann–Whitney test, n =3 (3 animals per group; each animal represented by 3 tissue sections). For ELISA in ( i , j , k and l ), statistical significance between 2 groups was determined by unpaired Student’s t -test (two-tailed) for two independent experiments with two donors/genotypes. * P <0.05 compared with WT. S, scar; T, tendon ends. Scale bars: 100 μ m

Article Snippet: Secreted biglycan and fibronectin were determined using mouse biglycan and fibronectin ELISA kits (Cloud-Clone Corp, Katy, TX, USA; and Aviva Systems Biology, San Diego, CA, USA; respectively) according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Tenomodulin is essential for prevention of adipocyte accumulation and fibrovascular scar formation during early tendon healing

doi: 10.1038/cddis.2017.510

Figure Lengend Snippet: The absence of Tnmd in TSPCs leads to significantly reduced cell migration and proliferation. ( a – d ) In vitro wound healing assays on collagen I and fibronectin showed that Tnmd −/− TSPC scratch closure was significantly slower compared with WT TSPCs. The borders of the scratches are outlined with yellow lines. ( e , f ) Forward migration index (FMI) plots showed that Tnmd −/− TSPCs were indeed less migratory than WT TSPCs. Upper arrows on each type of matrix show the start point while lower arrows the end point of example migratory cells. ( g–l ) Quantification of velocity, accumulated and Euclidean distances further validated Tnmd −/− migratory deficiency. ( m ) During 0, 4, 8 and 12 days of culture cell growth kinetics were estimated by DNA-based CyQUANT assay revealing that the proliferation of Tnmd −/− TSPCs was significantly lower than that of WT TSPCs. For quantification in ( c , d and m ), n =4 independent experiments per group. For quantifications in random migration, n =3 independent experiments per group (total of 70–80 tracks per genotype). Statistical significance was calculated using two-tailed non-parametric Mann–Whitney test. * P <0.05; ** P <0.01; *** P <0.001, compared with WT. Blue arrows, WT TSPCs; d, day; h, hour; Red arrows, Tnmd −/− TSPCs; Scale bars: 200 μ m

Article Snippet: Secreted biglycan and fibronectin were determined using mouse biglycan and fibronectin ELISA kits (Cloud-Clone Corp, Katy, TX, USA; and Aviva Systems Biology, San Diego, CA, USA; respectively) according to the manufacturer’s instructions.

Techniques: Migration, In Vitro, CyQUANT Assay, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in cardiovascular medicine

Article Title: Immune checkpoint inhibitor therapy increases systemic SDF-1, cardiac DAMPs Fibronectin-EDA, S100/Calgranulin, galectine-3, and NLRP3-MyD88-chemokine pathways.

doi: 10.3389/fcvm.2022.930797

Figure Lengend Snippet: FIGURE 8 Short-term ICI Therapy increases DAMPs, NLRP3 and MyD-88 mediated fibrosis and vascular inflammation. Short-term treatment with immune checkpoint inhibitors (ICIs) induced significant increases in DAMPs, galectine-3, cytokines and chemokines through NLRP3 and MyD88 pathways inducing myocardial hypertrophy, fibrosis and strong vascular inflammation.

Article Snippet: After centrifugation at 4◦C at 1,300 rpm for 10min, the supernatant of the cardiac homogenates were used to quantitative analysis of six biomarkers of cardiac damages and inflammation, such as: myeloid differentiation primary response 88 (MYd-88) expression (through mouse MyD88 ELISA Kit (My Biosource, San Diego, CA, detection range of 78–5,000 pg/ml; sensitivity: 46.9 pg/ml); NOD-, LRR- and pyrin domaincontaining protein 3 (NLRP-3) (through mouse NLRP3 ELISA Kit (OKEH05486, Aviva Systems Biology); Fibronectin-EDA, S100/Calgranulin and Galectine-3 [three DAMPs (quantified in cardiac tissues through selective quantitative assay); twelve cytokines and growth factors (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL10, IL-12, IL17-α, IFN-γ, TNF-α, G-CSF, GM-CSF) through a mouse cytokine Multiplex Assay kit (Qiagen, USA, pg/mg of heart tissue).

Techniques: