Journal: The Journal of Clinical Investigation

Article Title: PAI1 mediates fibroblast–mast cell interactions in skin fibrosis

doi: 10.1172/JCI99088

Figure Lengend Snippet: (A) Quantification of migrating mast cells into buffer or recombinant PAI1–containing media in a Transwell assay (n = 5). Quantification adherent mast cells on newborn dermal fibroblasts after (B) recombinant PAI1 treatment of fibroblast–mast cell cocultures (n = 4) and (C) pretreatment of fibroblasts with buffer, recombinant PAI1, and RGD peptide (n = 3). (D) Immunostaining for surface-bound vitronectin after treatment of fibroblasts with buffer or recombinant PAI1 (n = 3). Scale bar: 50 μm. (E) Quantification of adherent mast cells after pretreatment of fibroblasts with buffer or recombinant PAI1 in the absence or presence of FAK inhibitor (FAK inh) (n = 3). (F) Immunostaining of ICAM1 expression after recombinant PAI1 treatment of fibroblasts in the absence or presence of FAK inhibitor (n = 3). Scale bar: 50 μm. (G) Quantification of adherent mast cells after pretreatment of fibroblasts with buffer or recombinant PAI1 followed by incubation with ICAM1 inhibitor (ICAM1 inh) or LDV peptide, or with mast cells (MCs) preincubated with LDV peptide (n = 3). Data represent the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, by Student’s t test (A and B) and 1-way ANOVA followed by Tukey’s post hoc analysis (C, E, and G).

Article Snippet: The primary following antibodies were used at a dilution of 1:200: K5 (generated in-house); Pai1 (Abcam; ab66705); TENC (MilliporeSigma; ab19013); F4/80 (eBioscience; 14-4801-81); vitronectin (R&D Systems; MAB38751); ICAM1 (Thermo Fisher Scientific; 14-0541-81); α-SMA (MilliporeSigma; A2547 and Abcam; ab5694); vimentin (Abcam; AB92547); Ki67 (Abcam; ab16667); p-FAK (Thermo Fisher Scientific; 44624G); and FAK (Cell Signaling Technology; 3285S).

Techniques: Recombinant, Transwell Assay, Immunostaining, Expressing, Incubation

Journal: BMC Microbiology

Article Title: Nontypeable Haemophilus influenzae exploits the interaction between protein-E and vitronectin for the adherence and invasion to bronchial epithelial cells

doi: 10.1186/s12866-015-0600-8

Figure Lengend Snippet: NTHi binding of immobilized vitronectin in the absence or presence of heparin. Human plasma vitronectin was bound to glass-bottomed dishes. NTHi was incubated on the plate-bound vitronectin for 30 minutes, and then the dishes were washed three times. In some experiments, plate-bound vitronectin was pretreated with heparin before incubation with NTHi. NTHi was stained with LIVE/DEAD®. Live NTHi is stained green. BSA was used as a negative control. Representative fluorescent micrographs show that both NTHi strains (ATCC 19418 ( a ) and HUSM 0481 ( b )) attached to plate-bound vitronectin and that this attachment is blocked by heparin. White bars represent 10 μm. Summaries of the numbers of attached NTHi (per field at 1,000× magnification) are shown in ( c ) for ATCC 19418 and ( d ) for HUSM 0481. Error bars represent SEM in three independent experiments that gave similar results. ** p < 0.01

Article Snippet: After washing with PBS, 5.0 μg/ml of monoclonal antibody to human vitronectin (Takara, Otsu, Japan) was added and incubated for 60 minutes.

Techniques: Binding Assay, Incubation, Staining, Negative Control

Journal: BMC Microbiology

Article Title: Nontypeable Haemophilus influenzae exploits the interaction between protein-E and vitronectin for the adherence and invasion to bronchial epithelial cells

doi: 10.1186/s12866-015-0600-8

Figure Lengend Snippet: Binding of NTHi to immobilized vitronectin in the presence of protein-E peptide. Human plasma vitronectin was bound to glass-bottomed dishes. NTHi (HUSM 0481) was incubated on the plate-bound vitronectin for 30 minutes, and then the dishes were washed three times. In some experiments, plate-bound vitronectin was pretreated with protein-E peptide (PE 84–108 ) before incubation with NTHi. BSA was used as a negative control. NTHi was stained with LIVE/DEAD®, and viable NTHi is stained green. a Representative fluorescent micrographs of NTHi incubated on plate-bound vitronectin that was untreated or pretreated with PE 84–108 . White bars represent 10 μm. b The number of attached NTHi per field at 1,000 × magnification that was pretreated with increasing concentrations of PE 84–108 . Error bars represent SEM in three independent experiments that gave similar results. ** p < 0.01

Article Snippet: After washing with PBS, 5.0 μg/ml of monoclonal antibody to human vitronectin (Takara, Otsu, Japan) was added and incubated for 60 minutes.

Techniques: Binding Assay, Incubation, Negative Control, Staining

Journal: BMC Microbiology

Article Title: Nontypeable Haemophilus influenzae exploits the interaction between protein-E and vitronectin for the adherence and invasion to bronchial epithelial cells

doi: 10.1186/s12866-015-0600-8

Figure Lengend Snippet: A schema of the proposed mechanism by which NTHi penetrates into bronchial epithelial cell via protein-E and vitronectin. Vitronectin has three heparin-binding domains (HBDs), which interact with NTHi. Of those HBDs, the C-terminal HBD-3 corresponds to a protein-E binding region and interacts with PE 84–108 . This interaction is blocked by heparin or PE 84–108 peptide. Vitronectin also possesses a cell receptor binding site characterized by an Arg-Gly-Asp (RGD) sequence, which interacts with integrins on the bronchial epithelial cell surface. This protein-E-vitronectin axis seems to play a role in the adherence and penetration of NTHi into bronchial epithelial cells

Article Snippet: After washing with PBS, 5.0 μg/ml of monoclonal antibody to human vitronectin (Takara, Otsu, Japan) was added and incubated for 60 minutes.

Techniques: Binding Assay, Sequencing

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Proteomic analysis of FF samples revealed extracellular matrix (ECM) protein - vitronectin as the component responsible for FTE adhesion and spreading A. Experimental workflow of proteomics experiment. B. Venn diagram showing 14 common proteins identified between young and aged FF samples from proteomics analysis. Common proteins are listed with vitronectin (highlighted in red). C. Representative immunoblot for vitronectin expression in 3 young (Y1–Y3) and 3 aged (A1-A3) FF samples. FF samples (5 μg) and recombinant vitronectin protein (0.01 μg, 0.1 μg, 1 μg) diluted with lysis buffer were used for immunoblotting. Arrow represents the band analyzed for vitronectin. Ponceau staining was used for loading control.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Western Blot, Expressing, Recombinant, Lysis, Staining, Control

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Vitronectin in FF aids in FTE adhesion and spreading A. Representative brightfield images of FT190 cells seeded on ULA plates coated with FF sample (400 μl) and recombinant vitronectin protein (1 μg/well) for 4 h. Wells were washed with 1X PBS and FT190 cells were seeded on the coated plates. Images were acquired after 24 h. Scale bar = 200 μm. B. Cell proliferation was measured using an SRB assay to measure cell viability of FT190 cells on FF and vitronectin coated ULA plates. C. Representative Z stack images acquired by confocal microscopy of FT190 spheroids (labelled with Cell tracker 594) on NOF151 cells (labelled with Cell tracker 488) with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 200 μm. D. A maximum intensity projection re-construction of a three-dimensional FTE spheroid (labelled with Cell tracker 594) optical data stack over the surface of NOF151 cells (labelled with Cell tracker 488) treated with FF samples (Y1, A1) and vitronectin (1 μg). Spheroids were imaged with a 10× objective and re-constructed using the Imaris software. E. Representative images acquired by confocal microscopy showing a side and top projection of the FT190 spheroids on NOF151 cells with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 100 μm. F. Spheroid area (with and without FF/vitronectin) was determined by quantification of red fluorescent intensity using a polygon area measurement tool of the Imaris software.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Recombinant, Sulforhodamine B Assay, Confocal Microscopy, Construct, Software

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Primary antibodies.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: