mouse anti fibronectin primary antibody  (Developmental Studies Hybridoma Bank)

Journal: Development (Cambridge, England)

Article Title: Application and measurement of tissue-scale tension in avian epithelia in vivo to study multiscale mechanics and inter-germ layer coupling

doi: 10.1242/dev.204561

Figure Lengend Snippet: Mechanical coupling across germ layers. (A) Schematic of the embryonic region shown in B,C. Ventral view. A, anterior; ML, mediolateral; P, posterior. (B) Overlay of brightfield and fluorescent images prior to (top; applied strain E app =0) and after application of stretch to the endoderm (bottom; E app =0.6); endoderm nuclei are labeled by electroporation with pCAG-H2B-GFP (green). Magenta arrowheads and dashed yellow lines denote the positions of two nuclei and lateral somite boundaries, respectively. Scale bar: 100 µm. (C) Kymograph of stretch progressively applied to endoderm, illustrating concomitant deformation of somites over time. Scale bar: 100 µm. (D) Left: Schematic (top) and brightfield images (bottom) of the embryo following dorsal cuts to isolate ventral endoderm–somite interactions. Right: Comparison of Lagrangian strains in endoderm and mesoderm for control and dorsal cut embryos. ns, not significant ( P =0.52, Welch's t -test on the slopes of the strain transfer per sample). Circles represent the mean, error bars represent s.d. (E) Transverse view of fibronectin (FN; red) staining of the endoderm–somite interface of control (top) and Dispase-treated (bottom) embryos. DAPI was used to stain nuclei (gray). en, endoderm; no, notochord; so, somite. Scale bar: 10 µm. (F) Quantification of applied force versus in-plane Lagrangian strain in the endoderm for control and Dispase-treated embryos. (G) Relative stiffness quantified from the force-strain curves shown in F. * P =0.012 (Welch's t -test). (H) Comparison of Lagrangian strains in endoderm and mesoderm for control and Dispase-treated embryos. * P =0.012 (Welch's t -test on the slopes of the strain transfer per sample). Circles represent the mean, error bars represent s.d.

Article Snippet: Embryos were washed (PBS with 0.1% Triton X-100 was used for all washes) and left in blocking solution (10% heat inactivated goat serum) for 2 h, then incubated overnight at 4°C with mouse anti-fibronectin primary antibody (DSHB, B3/D6) diluted in PBS+0.1% Triton X-100 at 1:200.

Techniques: Labeling, Electroporation, Comparison, Control, Staining

Journal: Bioactive materials

Article Title: Multifunctionalized carbon-fiber-reinforced polyetheretherketone implant for rapid osseointegration under infected environment.

doi: 10.1016/j.bioactmat.2022.12.016

Figure Lengend Snippet: Fig. 1. Surface microstructures of the coatings. A) Surface microstructures of the CFRPEEK before and after Ti-PIII and hybrid PDA@ZnO NPs (or PDA@ZnO-EDN1) treatment. B) Energy-dispersive X-ray mapping of the Ti-PIII/PDA@ZnO-EDN1 surface. C) Water contact angles and photographs of water droplets on different samples. D) Vickers hardness of different samples. E) Ti and Zn ions concentration after immersion for different time. F) The release of EDN1 protein after immersion for different time. G) Increased fibronectin adsorption on modified surfaces. *p < 0.05, **p < 0.01 when compared with Ti-PIII; #p < 0.05, ##p < 0.01 when Ti-PIII/ PDA@ZnO-EDN1 compared with Ti-PIII/PDA@ZnO.

Article Snippet: Samples X. Wang et al. Bioactive Materials 24 (2023) 236–250 were incubated in a 24-well plate containing 1 mL Dulbecco’s modified Eagle medium (DMEM) with 10% fetal bovine serum (FBS) for 1 and 4 h at 37 ◦C and then treated with mouse monoclonal anti-fibronectin primary antibody (Santa Cruz Biotechnology, dilution, 1:50) and antimouse secondary antibody conjugated with horseradish peroxidase (HRP, Bio-Rad, MD, USA).

Techniques: Concentration Assay, Adsorption, Modification

Journal: Advanced Functional Materials

Article Title: A Bio‐Functional Wnt3a Gradient Microarray for Cell Niche Studies

doi: 10.1002/adfm.202301941

Figure Lengend Snippet: Figure 8. Maintenance of human hair dermal papilla cells (HHDPCs) on the Wnt3a gradient. A) Study design and workflow of characterizing the fibronectin meshwork deposited by the HHDPCs cultured on Wnt3a gradient. B) Representative immunofluorescence images of fibronectin deposited by the HHDPCs cultured on the NA gradient bound with bWnt3a (LOL = 12, 100 ng of input) i) and on the NA gradient only in the culture medium ii), for 24 h. Scale bar: 100 µm. Linear regression analysis of the relationship between the area of fibronectin matrix iii), the total length of fibronectin matrix iv), the percentage (%) of the high-density fibronectin matrix v) and the lacunarity of the fibronectin matrix vi) on the Wnt3a or NA gradient. Evaluation of β-catenin nuclear accumulation in HHDPCs cultured on the Wnt3a gradient by counting the number of cells positive in the nuclear β-catenin vii). n = 4 in two independent experiments. The differences of means among groups were analyzed by one-way ANOVA with Bonferroni’s post-hoc tests. *p < 0.05, **p < 0.01. C) Representative immunofluorescence images of ALP expression in the HHDPCs maintained on the NA gradient bound with bWnt3a (LOL = 12, 100 ng of input) for 24 h i) and the quantification of the area ii) and the level iii) of ALP expression. Scale bar: 100 µm. The differ- ences of means among groups were analyzed by Welch’s ANOVA with Dunnett’s T3 post-hoc tests. *p < 0.05, ***p = 0.0002, ****p < 0.0001. ns: not significant (p > 0.05). n = 9, 26, 32, and 30 cells in 18, 27, 36, and 45 mW group, respectively. D) Representative immunofluorescence images of BMP2 expression in the HHDPCs maintained on the NA gradient bound with bWnt3a (LOL = 12, 100 ng of input) for 24 h i) and the quantification of the area ii) and the level iii) of BMP2 expression. Scale bar: 100 µm. The differences of means among groups were analyzed by Welch’s ANOVA with Dunnett’s T3 post-hoc tests. ns: not significant (p > 0.05). n = 21, 20, 17, and 24 cells in 18, 27, 36, and 45 mW group, respectively. E) Representative immunofluo- rescence images of versican expression in the HHDPCs maintained on the NA gradient bound with bWnt3a (LOL = 12, 100 ng of input) for 24 h i) and the quantification of the area ii) and the level iii) of versican expression. Scale bar: 100 µm. The differences of means among groups were analyzed by Welch’s ANOVA with Dunnett’s T3 post-hoc tests. ns: not significant (p > 0.05). n = 12, 15, 39, and 38 cells in 18, 27, 36, and 45 mW group, respectively.

Article Snippet: Specifically, cells undergone fixation and permeabilization were incubated with mouse anti-fibronectin primary antibody (sc-8422, Santa Cruz Biotechnology, Dallas, TX, USA) (1:200) and rabbit anti-β-catenin primary antibody (ab16051, Abcam) (1:150) at 4 °C for overnight.

Techniques: Cell Culture, Immunofluorescence, Expressing