human fibronectin Search Results


fn 1918fn  (R&D Systems)
95
R&D Systems fn 1918fn
Fn 1918fn, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc04348401-212-13-17?v=R%26D+Systems
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fn 1918fn - by Bioz Stars, 2026-08
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human fibronectin  (R&D Systems)
93
R&D Systems human fibronectin
FIGURE 4. Stimulation of IVD cells with HTRA1-generated <t>fibronectin</t> fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.
Human Fibronectin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/10__1074_slash_jbc__m112__341032-51-0-8?v=R%26D+Systems
Average 93 stars, based on 1 article reviews
human fibronectin - by Bioz Stars, 2026-08
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fragment 3  (R&D Systems)
93
R&D Systems fragment 3
FIGURE 4. Stimulation of IVD cells with HTRA1-generated <t>fibronectin</t> fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.
Fragment 3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc05059943__13075_2016_1133_MOESM1_ESM-24-111-117?v=R%26D+Systems
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fragment 3 - by Bioz Stars, 2026-08
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acceptor bead solution antibody alphalisa acceptor human fibronectin r d systems  (R&D Systems)
95
R&D Systems acceptor bead solution antibody alphalisa acceptor human fibronectin r d systems
FIGURE 4. Stimulation of IVD cells with HTRA1-generated <t>fibronectin</t> fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.
Acceptor Bead Solution Antibody Alphalisa Acceptor Human Fibronectin R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc07822819__41598_2021_81253_MOESM1_ESM-57-315-323?v=R%26D+Systems
Average 95 stars, based on 1 article reviews
acceptor bead solution antibody alphalisa acceptor human fibronectin r d systems - by Bioz Stars, 2026-08
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recombinant human fibronectin  (R&D Systems)
93
R&D Systems recombinant human fibronectin
FIGURE 4. Stimulation of IVD cells with HTRA1-generated <t>fibronectin</t> fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.
Recombinant Human Fibronectin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc03320799-315-0-3?v=R%26D+Systems
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recombinant human fibronectin - by Bioz Stars, 2026-08
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fn1 neutralizing antibody fn1 ab  (R&D Systems)
93
R&D Systems fn1 neutralizing antibody fn1 ab
Transcriptome analysis revealed the central role of the ECM-PI3K pathway in pancreatic cancer metastasis. (A) Volcano plot of differentially expressed genes (DEGs), showing 248 DEGs between pancreatic cancer tissues (PANC) and adjacent normal tissues (CTR), with 153 genes upregulated and 95 genes downregulated. (B) Hierarchical clustering heatmap of the top 100 DEGs, demonstrating differences in gene expression patterns between PANC and CTR tissues. (C) Results of KEGG functional enrichment analysis, indicating that the DEGs are primarily involved in signaling pathways such as ECM-receptor interaction, Cytoskeleton in muscle cells, Focal adhesion, and P13K/AKT signaling pathway. (D) GO analysis results, showing significant enrichment of DEGs in processes related to adhesion and ECM. (E, F) CytoHubba analysis results, revealing <t>FN1</t> as a core hub gene in the P13K/AKT signaling pathway.
Fn1 Neutralizing Antibody Fn1 Ab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc12267018-75-0-7?v=R%26D+Systems
Average 93 stars, based on 1 article reviews
fn1 neutralizing antibody fn1 ab - by Bioz Stars, 2026-08
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sirna knockdown  (OriGene)
90
OriGene sirna knockdown
a , b Expression of GFP ( a ) and PD-L1 ( b ) on platelet surfaces after co-incubation with six FLAG-GFP- and PD-L1-GFP--transfected NSCLC cell lines (A549, NCI-H322, NCI-H522, NCI-H23, HOP-62, HOP-92) ( n = 3). c , d Ratio of GFP+ platelets/GFP+ tumor cells ( c ) and PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells ( d ) after co-incubation of platelets with transfected NSCLC cell lines ( n = 3). e Heat map of relative fibrinogen ( FBG ), tissue factor ( F3 ), fibronectin 1 ( FN1 ) and von Willebrand factor ( VWF ) mRNA levels in all tested NSCLC cell lines ( n = 3). f Relative mRNA level of FN1 in all tested NSCLC cell lines ( n = 3). g Correlation ratio of PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells and relative FN1 mRNA level ( n = 3). h Immunofluorescence images of platelet adhesion to NCI-H23 (upper left) and HOP-62 cells (upper right and lower) (fibronectin: green, platelets: red) ( n = 3). Upper scale bar 100 µm, lower 20 µm. i Left, Quantification of adhesive platelets after co-incubation with NCI-H23 and HOP-62. Quantified as CD61+ area in %/FoV ( n = 6 out of 3 independent experiments). Right, quantification of fibronectin covered area in %/FoV in NCI-H23 and HOP-62 cells ( n = 6 out of 3 independent experiments). j Correlation of platelet and fibronectin covered area in %/FoV. k Immunofluorescence images of PD-L1 and fibronectin expression in HOP-62 cells ( n = 2). Scale bar 20 µm. l Representative PLA with PD-L1 and fibronectin in HOP-62 cells. Scale bar left 20 µm, right 10 µm ( n = 1). m Representative PLA with PD-L1 and fibronectin in NCI-H23 and HOP-62 cell ( n = 3). Scale bar 10 µm. n PLA quantification of foci/cell in 119 NCI-H23 and 126 HOP-62 cells out of three biological replicates. o Western blot for PD-L1 and fibronectin in PD-L1-GFP-transfected HOP-62 cells after <t>siRNA</t> <t>knockdown</t> for fibronectin. Vinculin and α-Tubulin were used as loading controls ( n = 2). Presentation of full scan blots are provided in the Source data file. p Expression of PD-L1 on platelets after co-incubation with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). q Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). r Representative images of platelet adhesion to fibronectin-coated surface in the presence or absence of different platelet-blocking agents ( n = 3). Scale bar 20 µm. s , t Quantitative analysis of the platelet adhesion assay as platelet covered area/FoV in % (s) and platelets/FoV (t) ( n = 9 (small symbols) were analyzed out of three independent experiments (large symbols)). Horizontal lines represent mean. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. u Quantification of PD-L1 on platelets after co-incubation with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). v Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). a – d , f , p , q , u , v Data are mean ± SEM. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. g, j Correlation was determined by simple linear regression analysis. I, n Data are mean ± SEM. Statistical significance was calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.
Sirna Knockdown, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc08636618-344-1-20?v=OriGene
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sirna knockdown - by Bioz Stars, 2026-08
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quantikine elisa human fibronectin  (R&D Systems)
94
R&D Systems quantikine elisa human fibronectin
Dataset of conducted experiments with some descriptive statistics showing the sample size, median, range , and standard deviation
Quantikine Elisa Human Fibronectin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc07642058-95-23-29?v=R%26D+Systems
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quantikine elisa human fibronectin - by Bioz Stars, 2026-08
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fibronectin  (R&D Systems)
94
R&D Systems fibronectin
( A ) <t>Fibronectin</t> 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.
Fibronectin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc12416519-50-32-34?v=R%26D+Systems
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fibronectin - by Bioz Stars, 2026-08
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fibronectin duoset elisa  (R&D Systems)
94
R&D Systems fibronectin duoset elisa
Figure 1. αSMA gene expression and protein levels are reduced in TGFβ1-activated fibroblasts following ketotifen treatment. (A, B) HDFa and (C, D) WS1 fibroblasts were treated for 48 h with DMEM supplemented with 10% FBS as mock, 10 μM or 25 μM ketotifen, 10 ng/mL TGFβ1, or 10 ng/mL TGFβ1 with ketotifen added during the final 24 h. Gene expression of αSMA (ACTA2) was measured using RT-qPCR and normalized to housekeeping gene HPRT. HDFa cells were treated for 48 h under the same conditions and probed for αSMA protein by western blot. (E) A representative image of the blots is shown (left) and semi-quantitative assessments plotted (right) using GAPDH as loading control. Full-length blots are available in Supplementary Fig. 4. (F) COL1A1 was measured in HDFa cells treated with conditions using 25 μM ketotifen. Protein levels of pro-collagen 1α1 (G) and <t>fibronectin</t> (H) were determined by <t>ELISA</t> staining kits. Data shown as mean ± SEM. n = 3–6 per treatment condition. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. αSMA alpha-smooth muscle actin, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HDFa human dermal fibroblasts (adult), SEM standard error of the mean, TGFβ1 transforming growth factor-beta.
Fibronectin Duoset Elisa, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pm38528089-313-16-20?v=R%26D+Systems
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fibronectin duoset elisa - by Bioz Stars, 2026-08
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fibronectin mab1918  (R&D Systems)
94
R&D Systems fibronectin mab1918
Figure 1. αSMA gene expression and protein levels are reduced in TGFβ1-activated fibroblasts following ketotifen treatment. (A, B) HDFa and (C, D) WS1 fibroblasts were treated for 48 h with DMEM supplemented with 10% FBS as mock, 10 μM or 25 μM ketotifen, 10 ng/mL TGFβ1, or 10 ng/mL TGFβ1 with ketotifen added during the final 24 h. Gene expression of αSMA (ACTA2) was measured using RT-qPCR and normalized to housekeeping gene HPRT. HDFa cells were treated for 48 h under the same conditions and probed for αSMA protein by western blot. (E) A representative image of the blots is shown (left) and semi-quantitative assessments plotted (right) using GAPDH as loading control. Full-length blots are available in Supplementary Fig. 4. (F) COL1A1 was measured in HDFa cells treated with conditions using 25 μM ketotifen. Protein levels of pro-collagen 1α1 (G) and <t>fibronectin</t> (H) were determined by <t>ELISA</t> staining kits. Data shown as mean ± SEM. n = 3–6 per treatment condition. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. αSMA alpha-smooth muscle actin, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HDFa human dermal fibroblasts (adult), SEM standard error of the mean, TGFβ1 transforming growth factor-beta.
Fibronectin Mab1918, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+fibronectin/pmc03972339-149-16-20?v=R%26D+Systems
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fibronectin mab1918 - by Bioz Stars, 2026-08
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Image Search Results


FIGURE 4. Stimulation of IVD cells with HTRA1-generated fibronectin fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.

Journal: Journal of Biological Chemistry

Article Title: Detrimental Role for Human High Temperature Requirement Serine Protease A1 (HTRA1) in the Pathogenesis of Intervertebral Disc (IVD) Degeneration

doi: 10.1074/jbc.m112.341032

Figure Lengend Snippet: FIGURE 4. Stimulation of IVD cells with HTRA1-generated fibronectin fragments. A, concentrated protein supernatants (15 g) from IVD cells treated for 24 h without or with HTRA1mac (5 g/ml) or HTRA1macSA (5 g/ml) were subjected to immunoblotting using antibody Mab1935 specific for the fibronectincarboxyl-terminalheparin-bindingdomain(Cterminus)orMab1936specificforthefibronectinamino-terminalfibrin-andheparin-bindingdomain (N terminus). Fibronectin fragments containing the amino-terminal fibrin- and heparin-binding domain are identified by the closed arrowhead. B, purified human plasma-derived fibronectin (Fn) was incubated with HTRA1mac or HTRA1macSA at equimolar concentrations in TBS, pH 8.5, for 16 h at 37 °C, and samples were loaded onto a 4–15% gradient gel and stained with Coomassie Blue. Fibronectin and recombinant HTRA1 alone were also loaded and served as controls. C, an equimolar concentration of human plasma-derived fibronectin and HTRA1mac were incubated for 16 h, and fibronectin fragments were visualized by Western blot analysis using the antibodies described in A. D, equimolar concentrations of fibronectin (20 g) and HTRA1mac (5 g) were incubated for 16 h, and fibronectin fragments were purified by affinity chromatography. IVD cells were incubated with purified HTRA1-digested fibronectin (FnHTRA1mac) for 24 h, and expression levels of MMP1, MMP3, and MMP13 mRNA were determined by qRT-PCR and the -fold change as compared with untreatedcontrolswasdeterminedusingthe2CTmethod.Additionalcultureswereincubatedwitheitheraffinity-purifiedTris-bufferedsaline,pH7.6(TBS), fibronectin (Fn), or HTRA1 (HTRA1mac) or left untreated (Control). Data are representative of two separate experiments performed using IVD cells from two patients. Shown are results of triplicate determinations S.D. *, p 0.01, as determined by one-way ANOVA.

Article Snippet: Materials—Human fibronectin and rabbit IgG were purchased from R & D Systems (Abingdon, UK).

Techniques: Generated, Western Blot, Binding Assay, Purification, Clinical Proteomics, Derivative Assay, Incubation, Staining, Recombinant, Concentration Assay, Affinity Chromatography, Expressing, Quantitative RT-PCR, Control

FIGURE 5. Detection of fibronectin fragments in degenerated IVD tissue. A, fibronectin (FN) mRNA levels in intact IVD tissue samples from patients (n 36) with varying degrees of IVD degeneration were determined by qRT-PCR and presented as 2CT S.E. (error bars). B, correlation study between FN and HTRA1 mRNA levels (2CT) in patient IVD tissue samples (n 36). R2, square of correlation coefficient; p 0.01 as determined from Pearson’s correlation coefficient. C, protein extracts from patient IVD tissues (n 12) were loaded onto a 12% SDS-polyacrylamide gel, and immunoblotting was performed using a monoclonal antibody (Mab1936) specific for the amino-terminal fibrin- and heparin-binding domain. D, the PVDF membrane used in C was stained with Coomassie Blue in order to confirm equal protein loading. Lane 1, HTRA1-digested human plasma-derived fibronectin; lanes 2–4, non-degenerated (ND) discs; lanes 5–7, mildly degenerated discs; lanes 8–10, moderately degenerated discs; lanes 11–13, severely degenerated discs.

Journal: Journal of Biological Chemistry

Article Title: Detrimental Role for Human High Temperature Requirement Serine Protease A1 (HTRA1) in the Pathogenesis of Intervertebral Disc (IVD) Degeneration

doi: 10.1074/jbc.m112.341032

Figure Lengend Snippet: FIGURE 5. Detection of fibronectin fragments in degenerated IVD tissue. A, fibronectin (FN) mRNA levels in intact IVD tissue samples from patients (n 36) with varying degrees of IVD degeneration were determined by qRT-PCR and presented as 2CT S.E. (error bars). B, correlation study between FN and HTRA1 mRNA levels (2CT) in patient IVD tissue samples (n 36). R2, square of correlation coefficient; p 0.01 as determined from Pearson’s correlation coefficient. C, protein extracts from patient IVD tissues (n 12) were loaded onto a 12% SDS-polyacrylamide gel, and immunoblotting was performed using a monoclonal antibody (Mab1936) specific for the amino-terminal fibrin- and heparin-binding domain. D, the PVDF membrane used in C was stained with Coomassie Blue in order to confirm equal protein loading. Lane 1, HTRA1-digested human plasma-derived fibronectin; lanes 2–4, non-degenerated (ND) discs; lanes 5–7, mildly degenerated discs; lanes 8–10, moderately degenerated discs; lanes 11–13, severely degenerated discs.

Article Snippet: Materials—Human fibronectin and rabbit IgG were purchased from R & D Systems (Abingdon, UK).

Techniques: Quantitative RT-PCR, Western Blot, Binding Assay, Membrane, Staining, Clinical Proteomics, Derivative Assay

FIGURE 6. A theoretical model for the role of HTRA1 in IVD degeneration. Based on our findings, we propose that HTRA1 accumulates in IVD tissue undergoing degeneration and stimulates MMP production by resident cells in a predominantly protease-dependent manner, via activation of the MEK pathway. Furthermore, we suggest that the stimulatory effects of HTRA1 on IVD cells are mediated indirectly through its ability to generate fibronectin fragments, although other routes of cellular activation cannot be ruled out. IDD, intervertebral disc degeneration.

Journal: Journal of Biological Chemistry

Article Title: Detrimental Role for Human High Temperature Requirement Serine Protease A1 (HTRA1) in the Pathogenesis of Intervertebral Disc (IVD) Degeneration

doi: 10.1074/jbc.m112.341032

Figure Lengend Snippet: FIGURE 6. A theoretical model for the role of HTRA1 in IVD degeneration. Based on our findings, we propose that HTRA1 accumulates in IVD tissue undergoing degeneration and stimulates MMP production by resident cells in a predominantly protease-dependent manner, via activation of the MEK pathway. Furthermore, we suggest that the stimulatory effects of HTRA1 on IVD cells are mediated indirectly through its ability to generate fibronectin fragments, although other routes of cellular activation cannot be ruled out. IDD, intervertebral disc degeneration.

Article Snippet: Materials—Human fibronectin and rabbit IgG were purchased from R & D Systems (Abingdon, UK).

Techniques: Activation Assay

Transcriptome analysis revealed the central role of the ECM-PI3K pathway in pancreatic cancer metastasis. (A) Volcano plot of differentially expressed genes (DEGs), showing 248 DEGs between pancreatic cancer tissues (PANC) and adjacent normal tissues (CTR), with 153 genes upregulated and 95 genes downregulated. (B) Hierarchical clustering heatmap of the top 100 DEGs, demonstrating differences in gene expression patterns between PANC and CTR tissues. (C) Results of KEGG functional enrichment analysis, indicating that the DEGs are primarily involved in signaling pathways such as ECM-receptor interaction, Cytoskeleton in muscle cells, Focal adhesion, and P13K/AKT signaling pathway. (D) GO analysis results, showing significant enrichment of DEGs in processes related to adhesion and ECM. (E, F) CytoHubba analysis results, revealing FN1 as a core hub gene in the P13K/AKT signaling pathway.

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Transcriptome analysis revealed the central role of the ECM-PI3K pathway in pancreatic cancer metastasis. (A) Volcano plot of differentially expressed genes (DEGs), showing 248 DEGs between pancreatic cancer tissues (PANC) and adjacent normal tissues (CTR), with 153 genes upregulated and 95 genes downregulated. (B) Hierarchical clustering heatmap of the top 100 DEGs, demonstrating differences in gene expression patterns between PANC and CTR tissues. (C) Results of KEGG functional enrichment analysis, indicating that the DEGs are primarily involved in signaling pathways such as ECM-receptor interaction, Cytoskeleton in muscle cells, Focal adhesion, and P13K/AKT signaling pathway. (D) GO analysis results, showing significant enrichment of DEGs in processes related to adhesion and ECM. (E, F) CytoHubba analysis results, revealing FN1 as a core hub gene in the P13K/AKT signaling pathway.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Gene Expression, Functional Assay, Protein-Protein interactions

Single-cell sequencing identifies specific expression patterns of PI3K-related genes including FN1 in fibroblasts of metastatic samples. (A) UMAP plot displaying the sub-cell types in the primary pancreatic cancer (stiu) and liver metastasis (meta) samples from the single-cell RNA sequencing dataset GSE156405 . (B) UMAP plot showing the expression patterns of different groups in primary pancreatic cancer (stiu) and liver metastasis (meta) samples. (C) Cell proportion plot illustrating the distribution of different cell types in stiu and meta samples. (D) Bubble plot representing the relationship between different sub-cell types and characteristic gene expression. The color of the bubbles ranges from white to blue, representing gene expression percentages of 0%, 25%, 50%, and 75%, respectively. The size of the bubbles indicates the average expression level, ranging from a minimum to a maximum representing average expression values from 0 to 2. (E) KEGG enrichment analysis of DEGs in alveolar epithelial cells. (F) KEGG enrichment analysis of DEGs in fibroblasts. (G) Violin plot showing the expression patterns of PI3K-related genes (FN1, THBS2, COL1A1, COL1A2, and COL6A3) in different cell types and groups in fibroblasts.

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Single-cell sequencing identifies specific expression patterns of PI3K-related genes including FN1 in fibroblasts of metastatic samples. (A) UMAP plot displaying the sub-cell types in the primary pancreatic cancer (stiu) and liver metastasis (meta) samples from the single-cell RNA sequencing dataset GSE156405 . (B) UMAP plot showing the expression patterns of different groups in primary pancreatic cancer (stiu) and liver metastasis (meta) samples. (C) Cell proportion plot illustrating the distribution of different cell types in stiu and meta samples. (D) Bubble plot representing the relationship between different sub-cell types and characteristic gene expression. The color of the bubbles ranges from white to blue, representing gene expression percentages of 0%, 25%, 50%, and 75%, respectively. The size of the bubbles indicates the average expression level, ranging from a minimum to a maximum representing average expression values from 0 to 2. (E) KEGG enrichment analysis of DEGs in alveolar epithelial cells. (F) KEGG enrichment analysis of DEGs in fibroblasts. (G) Violin plot showing the expression patterns of PI3K-related genes (FN1, THBS2, COL1A1, COL1A2, and COL6A3) in different cell types and groups in fibroblasts.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Sequencing, Expressing, RNA Sequencing, Gene Expression

Core mechanisms of CAFs in pancreatic cancer metastasis. (A) UMAP plot displaying fibroblast subpopulations by cluster. (B) UMAP plot showing the distribution of fibroblast subpopulations by group. (C) Proportion plot illustrating the cell proportions by cell subpopulation and group. (D) Identification of the CAFs subpopulation among fibroblast subpopulations. (E) UMAP visualization analysis, displaying the expression patterns of FN1, THBS2, COL1A1, COL1A2, and COL6A3 in the stiu and meta groups.

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Core mechanisms of CAFs in pancreatic cancer metastasis. (A) UMAP plot displaying fibroblast subpopulations by cluster. (B) UMAP plot showing the distribution of fibroblast subpopulations by group. (C) Proportion plot illustrating the cell proportions by cell subpopulation and group. (D) Identification of the CAFs subpopulation among fibroblast subpopulations. (E) UMAP visualization analysis, displaying the expression patterns of FN1, THBS2, COL1A1, COL1A2, and COL6A3 in the stiu and meta groups.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Expressing

CAF-derived FN1 promotes invasion and migration of pancreatic cancer cells. (A) Schematic diagram of the CAFs-PANC1 co-culture model. (B) ELISA results showing changes in IL-6, IL-8, and MMP2 levels secreted by CAFs before and after TGF-β induction. (C) ELISA results demonstrating changes in FN1 levels secreted by CAFs before and after treatment with FN1 neutralizing antibody (FN1-Ab). (D) Transwell migration assay showing changes in the number of migrated cells in the FN1-Ab group compared to the control group. (E) Statistics of migration rates from the Transwell migration assay. (F) Transwell invasion assay showing changes in the number of invading cells in the FN1-Ab group compared to the control group. (G) Statistics of invasion rates from the Transwell invasion assay.

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: CAF-derived FN1 promotes invasion and migration of pancreatic cancer cells. (A) Schematic diagram of the CAFs-PANC1 co-culture model. (B) ELISA results showing changes in IL-6, IL-8, and MMP2 levels secreted by CAFs before and after TGF-β induction. (C) ELISA results demonstrating changes in FN1 levels secreted by CAFs before and after treatment with FN1 neutralizing antibody (FN1-Ab). (D) Transwell migration assay showing changes in the number of migrated cells in the FN1-Ab group compared to the control group. (E) Statistics of migration rates from the Transwell migration assay. (F) Transwell invasion assay showing changes in the number of invading cells in the FN1-Ab group compared to the control group. (G) Statistics of invasion rates from the Transwell invasion assay.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Derivative Assay, Migration, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Transwell Migration Assay, Control, Transwell Invasion Assay

The FN1-ITG-PI3K/AKT axis promotes invasion and migration of pancreatic cancer cells. (A, B) GEPIA database analysis revealing a significant positive correlation between FN1 expression and the expression of key genes in the PI3K/AKT pathway (PIK3CA, AKT1). (C, D) Western blot results showing changes in phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab group. The Western blot results demonstrated the changes in the phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab groups of PANC-1 and BXPC-3 cells. (E, F) Statistical data corresponding to (C, D) . (G, H) mRNA expression changes of integrin genes (ITGA2, ITGB4, ITGA3) before and after treatment with an integrin inhibitor (ITG-Inh). The changes in mRNA expression of integrin genes ITGA2 , ITGB4 , and ITGA3 in PANC-1 and BXPC-3 cells before and after treatment with the integrin inhibitor ITG-Inh. (I, J) Western blot results demonstrating changes in protein levels of p-AKT and p-PI3K in the ITG-Inh group compared to the control group (ITG-Con). The Western blot results indicated that in PANC-1 and BXPC-3 cells, compared with the control group (ITG-CON), the protein levels of p-AKT and p-PI3K in the ITG-Inh group were altered. (K, L) Statistical data corresponding to (I, J) . (M, N) Changes in invasion ability after combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh). The changes in invasive capacity following the combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh) in PANC-1 and BXPC-3 cells. (O, P) Statistical data corresponding to (M, N) .

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: The FN1-ITG-PI3K/AKT axis promotes invasion and migration of pancreatic cancer cells. (A, B) GEPIA database analysis revealing a significant positive correlation between FN1 expression and the expression of key genes in the PI3K/AKT pathway (PIK3CA, AKT1). (C, D) Western blot results showing changes in phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab group. The Western blot results demonstrated the changes in the phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab groups of PANC-1 and BXPC-3 cells. (E, F) Statistical data corresponding to (C, D) . (G, H) mRNA expression changes of integrin genes (ITGA2, ITGB4, ITGA3) before and after treatment with an integrin inhibitor (ITG-Inh). The changes in mRNA expression of integrin genes ITGA2 , ITGB4 , and ITGA3 in PANC-1 and BXPC-3 cells before and after treatment with the integrin inhibitor ITG-Inh. (I, J) Western blot results demonstrating changes in protein levels of p-AKT and p-PI3K in the ITG-Inh group compared to the control group (ITG-Con). The Western blot results indicated that in PANC-1 and BXPC-3 cells, compared with the control group (ITG-CON), the protein levels of p-AKT and p-PI3K in the ITG-Inh group were altered. (K, L) Statistical data corresponding to (I, J) . (M, N) Changes in invasion ability after combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh). The changes in invasive capacity following the combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh) in PANC-1 and BXPC-3 cells. (O, P) Statistical data corresponding to (M, N) .

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Migration, Expressing, Western Blot, Phospho-proteomics, Control, Inhibition

High FN1 expression is associated with poor prognosis and an immunosuppressive microenvironment. (A) Clinical data analysis showing the relationship between FN1 expression and patient survival. (B) TIMER2.0 displaying the correlation between FN1 expression and M2 macrophage infiltration in pancreatic cancer (PAAD). (C) TIMER2.0 showing the correlation between FN1 expression and Treg cell infiltration in PAAD. (D) Mechanism diagram illustrating how FN1 activates the PI3K/AKT pathway by binding to integrin receptors, thereby promoting the invasion and metastasis of pancreatic cancer cells.

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: High FN1 expression is associated with poor prognosis and an immunosuppressive microenvironment. (A) Clinical data analysis showing the relationship between FN1 expression and patient survival. (B) TIMER2.0 displaying the correlation between FN1 expression and M2 macrophage infiltration in pancreatic cancer (PAAD). (C) TIMER2.0 showing the correlation between FN1 expression and Treg cell infiltration in PAAD. (D) Mechanism diagram illustrating how FN1 activates the PI3K/AKT pathway by binding to integrin receptors, thereby promoting the invasion and metastasis of pancreatic cancer cells.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Expressing, Binding Assay

a , b Expression of GFP ( a ) and PD-L1 ( b ) on platelet surfaces after co-incubation with six FLAG-GFP- and PD-L1-GFP--transfected NSCLC cell lines (A549, NCI-H322, NCI-H522, NCI-H23, HOP-62, HOP-92) ( n = 3). c , d Ratio of GFP+ platelets/GFP+ tumor cells ( c ) and PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells ( d ) after co-incubation of platelets with transfected NSCLC cell lines ( n = 3). e Heat map of relative fibrinogen ( FBG ), tissue factor ( F3 ), fibronectin 1 ( FN1 ) and von Willebrand factor ( VWF ) mRNA levels in all tested NSCLC cell lines ( n = 3). f Relative mRNA level of FN1 in all tested NSCLC cell lines ( n = 3). g Correlation ratio of PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells and relative FN1 mRNA level ( n = 3). h Immunofluorescence images of platelet adhesion to NCI-H23 (upper left) and HOP-62 cells (upper right and lower) (fibronectin: green, platelets: red) ( n = 3). Upper scale bar 100 µm, lower 20 µm. i Left, Quantification of adhesive platelets after co-incubation with NCI-H23 and HOP-62. Quantified as CD61+ area in %/FoV ( n = 6 out of 3 independent experiments). Right, quantification of fibronectin covered area in %/FoV in NCI-H23 and HOP-62 cells ( n = 6 out of 3 independent experiments). j Correlation of platelet and fibronectin covered area in %/FoV. k Immunofluorescence images of PD-L1 and fibronectin expression in HOP-62 cells ( n = 2). Scale bar 20 µm. l Representative PLA with PD-L1 and fibronectin in HOP-62 cells. Scale bar left 20 µm, right 10 µm ( n = 1). m Representative PLA with PD-L1 and fibronectin in NCI-H23 and HOP-62 cell ( n = 3). Scale bar 10 µm. n PLA quantification of foci/cell in 119 NCI-H23 and 126 HOP-62 cells out of three biological replicates. o Western blot for PD-L1 and fibronectin in PD-L1-GFP-transfected HOP-62 cells after siRNA knockdown for fibronectin. Vinculin and α-Tubulin were used as loading controls ( n = 2). Presentation of full scan blots are provided in the Source data file. p Expression of PD-L1 on platelets after co-incubation with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). q Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). r Representative images of platelet adhesion to fibronectin-coated surface in the presence or absence of different platelet-blocking agents ( n = 3). Scale bar 20 µm. s , t Quantitative analysis of the platelet adhesion assay as platelet covered area/FoV in % (s) and platelets/FoV (t) ( n = 9 (small symbols) were analyzed out of three independent experiments (large symbols)). Horizontal lines represent mean. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. u Quantification of PD-L1 on platelets after co-incubation with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). v Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). a – d , f , p , q , u , v Data are mean ± SEM. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. g, j Correlation was determined by simple linear regression analysis. I, n Data are mean ± SEM. Statistical significance was calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Platelet PD-L1 reflects collective intratumoral PD-L1 expression and predicts immunotherapy response in non-small cell lung cancer

doi: 10.1038/s41467-021-27303-7

Figure Lengend Snippet: a , b Expression of GFP ( a ) and PD-L1 ( b ) on platelet surfaces after co-incubation with six FLAG-GFP- and PD-L1-GFP--transfected NSCLC cell lines (A549, NCI-H322, NCI-H522, NCI-H23, HOP-62, HOP-92) ( n = 3). c , d Ratio of GFP+ platelets/GFP+ tumor cells ( c ) and PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells ( d ) after co-incubation of platelets with transfected NSCLC cell lines ( n = 3). e Heat map of relative fibrinogen ( FBG ), tissue factor ( F3 ), fibronectin 1 ( FN1 ) and von Willebrand factor ( VWF ) mRNA levels in all tested NSCLC cell lines ( n = 3). f Relative mRNA level of FN1 in all tested NSCLC cell lines ( n = 3). g Correlation ratio of PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells and relative FN1 mRNA level ( n = 3). h Immunofluorescence images of platelet adhesion to NCI-H23 (upper left) and HOP-62 cells (upper right and lower) (fibronectin: green, platelets: red) ( n = 3). Upper scale bar 100 µm, lower 20 µm. i Left, Quantification of adhesive platelets after co-incubation with NCI-H23 and HOP-62. Quantified as CD61+ area in %/FoV ( n = 6 out of 3 independent experiments). Right, quantification of fibronectin covered area in %/FoV in NCI-H23 and HOP-62 cells ( n = 6 out of 3 independent experiments). j Correlation of platelet and fibronectin covered area in %/FoV. k Immunofluorescence images of PD-L1 and fibronectin expression in HOP-62 cells ( n = 2). Scale bar 20 µm. l Representative PLA with PD-L1 and fibronectin in HOP-62 cells. Scale bar left 20 µm, right 10 µm ( n = 1). m Representative PLA with PD-L1 and fibronectin in NCI-H23 and HOP-62 cell ( n = 3). Scale bar 10 µm. n PLA quantification of foci/cell in 119 NCI-H23 and 126 HOP-62 cells out of three biological replicates. o Western blot for PD-L1 and fibronectin in PD-L1-GFP-transfected HOP-62 cells after siRNA knockdown for fibronectin. Vinculin and α-Tubulin were used as loading controls ( n = 2). Presentation of full scan blots are provided in the Source data file. p Expression of PD-L1 on platelets after co-incubation with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). q Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). r Representative images of platelet adhesion to fibronectin-coated surface in the presence or absence of different platelet-blocking agents ( n = 3). Scale bar 20 µm. s , t Quantitative analysis of the platelet adhesion assay as platelet covered area/FoV in % (s) and platelets/FoV (t) ( n = 9 (small symbols) were analyzed out of three independent experiments (large symbols)). Horizontal lines represent mean. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. u Quantification of PD-L1 on platelets after co-incubation with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). v Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). a – d , f , p , q , u , v Data are mean ± SEM. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. g, j Correlation was determined by simple linear regression analysis. I, n Data are mean ± SEM. Statistical significance was calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.

Article Snippet: For siRNA knockdown of Fibronectin, Lipofectamine TM 3000 and 100 pmol of human (FN1) siRNA Oligo Duplex (Locus ID 2335) (Origene, SR320193) was used.

Techniques: Expressing, Incubation, Transfection, Immunofluorescence, Adhesive, Western Blot, Knockdown, Blocking Assay, Cell Adhesion Assay, Two Tailed Test

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

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

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

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

( 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.

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

Figure 1. αSMA gene expression and protein levels are reduced in TGFβ1-activated fibroblasts following ketotifen treatment. (A, B) HDFa and (C, D) WS1 fibroblasts were treated for 48 h with DMEM supplemented with 10% FBS as mock, 10 μM or 25 μM ketotifen, 10 ng/mL TGFβ1, or 10 ng/mL TGFβ1 with ketotifen added during the final 24 h. Gene expression of αSMA (ACTA2) was measured using RT-qPCR and normalized to housekeeping gene HPRT. HDFa cells were treated for 48 h under the same conditions and probed for αSMA protein by western blot. (E) A representative image of the blots is shown (left) and semi-quantitative assessments plotted (right) using GAPDH as loading control. Full-length blots are available in Supplementary Fig. 4. (F) COL1A1 was measured in HDFa cells treated with conditions using 25 μM ketotifen. Protein levels of pro-collagen 1α1 (G) and fibronectin (H) were determined by ELISA staining kits. Data shown as mean ± SEM. n = 3–6 per treatment condition. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. αSMA alpha-smooth muscle actin, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HDFa human dermal fibroblasts (adult), SEM standard error of the mean, TGFβ1 transforming growth factor-beta.

Journal: Scientific reports

Article Title: Ketotifen directly modifies the fibrotic response of human skin fibroblasts.

doi: 10.1038/s41598-024-57776-7

Figure Lengend Snippet: Figure 1. αSMA gene expression and protein levels are reduced in TGFβ1-activated fibroblasts following ketotifen treatment. (A, B) HDFa and (C, D) WS1 fibroblasts were treated for 48 h with DMEM supplemented with 10% FBS as mock, 10 μM or 25 μM ketotifen, 10 ng/mL TGFβ1, or 10 ng/mL TGFβ1 with ketotifen added during the final 24 h. Gene expression of αSMA (ACTA2) was measured using RT-qPCR and normalized to housekeeping gene HPRT. HDFa cells were treated for 48 h under the same conditions and probed for αSMA protein by western blot. (E) A representative image of the blots is shown (left) and semi-quantitative assessments plotted (right) using GAPDH as loading control. Full-length blots are available in Supplementary Fig. 4. (F) COL1A1 was measured in HDFa cells treated with conditions using 25 μM ketotifen. Protein levels of pro-collagen 1α1 (G) and fibronectin (H) were determined by ELISA staining kits. Data shown as mean ± SEM. n = 3–6 per treatment condition. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. αSMA alpha-smooth muscle actin, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HDFa human dermal fibroblasts (adult), SEM standard error of the mean, TGFβ1 transforming growth factor-beta.

Article Snippet: Antibodies used for protein concentration measurements were from pro-collagen 1α1 DuoSet ELISA (DY6220-05, R&D Systems) and fibronectin DuoSet ELISA (DY1918-05, R&D Systems) staining kits.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Staining