integrin beta 4 β4  (Proteintech)

Journal: International Journal of Nanomedicine

Article Title: Highly Ordered Nanotube-Like Microstructure on Titanium Dental Implant Surface Fabricated via Anodization Enhanced Cell Adhesion and Migration of Human Gingival Fibroblasts

doi: 10.2147/IJN.S448743

Figure Lengend Snippet: Gene and protein expression of adhesion molecules. The presence of nanotubes regulated adhesion-related gene expression of fibroblasts ( A ). It was noteworthy that integrin β1 (ITGB1) was upregulated, in all nanotube groups at all timepoints compared with RT (p<0.05). Fibronectin (FN) was upregulated in TNT-30 and 40 at 6 hr and 2 d of cell culture compared with RT and TNT-50 (p<0.05). Integrin α6 (ITGA6) was upregulated at 6 hr in TNT-30 and TNT-40 (p<0.05). At later stage of cell adhesion (12 hr and 2 d), Integrin β4 (ITGB4) was just upregulated in TNT30 and 40 (p<0.05). Intercellular cell adhesion molecule-1 (ICAM-1) was significantly upregulated in TNT-30 and 40 at 6 hr and 12 hr of cell culture compared with RT and TNT-50 (p<0.05). Gene expression of type I collagen was upregulated in TNT-30 and 40 at all timepoints of cell culture, compared with RT (p<0.05). Protein expression of Integrin β1/β4/α6 and fibronectin was evaluated via Western blot and the results of protein expression of adhesive molecule demonstrated that compared with the raw Ti, the presence of nanotubes on the surface exerted a positive role in regulating cell adhesion, especially the expression of integrin β1 and fibronectin at all the timepoints of cell culture ( B and C ). The presence of nanotubes surface increased the expression of integrin α6 at the initial 6th hour and integrin β4 at the 12th hour. At other timepoints, protein expression of integrin α6 and β4 was not significantly altered between groups. (*Represented p<0.05 compared with RT, # Represented p<0.05 compared with TNT-50).

Article Snippet: After blocking with 10% skimmed milk, the membrane was probed overnight at 4°C using specific primary antibodies including integrin β1 (26918-1-AP,1:1000, ProteinTech, China), Fibronectin (15613-1-AP, 1:1000, ProteinTech, China), Integrin α6 (27189-1-AP, 1:1000, ProteinTech, China), and Integrin β4 (21738-1-AP, 1:1000, ProteinTech, China), followed by incubation with a horseradish peroxidase–conjugated secondary antibody, HRP-goat anti-mouse or HRP-goat anti-rabbit, for 1 hr at room temperature.

Techniques: Expressing, Gene Expression, Cell Culture, Western Blot, Adhesive

Journal: Cancers

Article Title: Human Breast Extracellular Matrix Microstructures and Protein Hydrogel 3D Cultures of Mammary Epithelial Cells

doi: 10.3390/cancers13225857

Figure Lengend Snippet: HB-TMG support of human breast epithelial cell 3D growth. ( a ) MCF10A cell suspending growth within HB-TMG in the presence or absence of E2 or/and IGF-I stimulation. Arrows, cell spheres. Scale bars, 50 μm. ( b ) Quantification of the volumes (vol) of the Day 12 MCF10A cell 3D aggregates shown in ( a ). * p < 0.05; ** p < 0.01. ( c ) IF staining of normal breast or breast cancer epithelial cell culture on 2D surfaces or 3D HB-TMG. Arrows, cell protrusions. Red, phalloidin; green, Col I; blue, nucleus Hoechst staining. Scale bars, 5 μm. ( d ) Quantification of cell protrusions in 2D cultures. * p < 0.05; ** p < 0.01. ( e ) Quantification of cell protrusions in 3D cultures. ( f ) IF staining of acini formation of normal breast or breast cancer epithelial cells. Red, β4 integrin; green, E-cadherin; blue, nucleus Hoechst staining. Scale bars: c-i, c-ii, and c-iv, 20 μm; c-iii, 50 μm.

Article Snippet: The cells were fixed with paraformaldehyde, permeabilized, and incubated with primary antibodies against E-cadherin (Thermo Fisher Scientific, #701134) and β4 integrin (R&D Systems, Minneapolis, MN, USA; #MAB4060), followed by fluorophore-conjugated secondary antibodies and Hoechst staining as described above, imaged for acinar structures under a fluorescent microscope.

Techniques: Staining, Cell Culture