rabbit anti human sema4d  (Bioss)

Journal: Dentistry Journal

Article Title: Semaphorin 4D Induces Vasculogenic Differentiation of Dental Pulp Stem Cells

doi: 10.3390/dj11070160

Figure Lengend Snippet: Semaphorin 4D and Plexin B1 expression in the dental pulp. ( A ) Western blot analysis shows the protein expression of SEMA4D and Plexin B1 in lysates from DPSC, SHED cells and human dental pulp tissue. Primary human dermal microvascular endothelial cells (HDMEC) were used as controls. ( B ) Immunofluorescence staining of pulp tissues for SEMA4D or isotype-control IgG. ( C ) Immunofluorescence staining of pulp tissues for Plexin B1 or isotype-control IgG. Top images ( B , C ) depict pulp tissues from human non-carious third molars, whereas bottom images show tissues from tooth slice scaffolds seeded with DPSC cells and 3 weeks after transplantation into SCID mice (images taken at 200× magnification, scale bars: 50 μm).

Article Snippet: After antigen retrieval, tissue sections were incubated overnight at 4 °C with rabbit anti-human SEMA4D (Bioss Inc.), mouse anti-human Plexin B1 (Santa Cruz Biotechnology Inc.) or non-specific isotype-matched IgG that was used as a negative control.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Transplantation Assay

Journal: Dentistry Journal

Article Title: Semaphorin 4D Induces Vasculogenic Differentiation of Dental Pulp Stem Cells

doi: 10.3390/dj11070160

Figure Lengend Snippet: Semaphorin 4D induces expression of endothelial cell markers in dental pulp stem cells. Western blot analysis of DPSC and SHED treated with 0, 25, 50, or 100 ng/mL of rhSEMA4D for 7 days depicts the expression of VEGFR2, CD31, and Tie2. Primary human dermal microvascular endothelial cells (HDMEC) were used as controls.

Article Snippet: After antigen retrieval, tissue sections were incubated overnight at 4 °C with rabbit anti-human SEMA4D (Bioss Inc.), mouse anti-human Plexin B1 (Santa Cruz Biotechnology Inc.) or non-specific isotype-matched IgG that was used as a negative control.

Techniques: Expressing, Western Blot

Journal: Dentistry Journal

Article Title: Semaphorin 4D Induces Vasculogenic Differentiation of Dental Pulp Stem Cells

doi: 10.3390/dj11070160

Figure Lengend Snippet: Semaphorin 4D induces DPSC cells to differentiate into sprout-like structures. ( A ) Photomicrographs of DPSC (×100 magnification) seeded in a 12-well plate (1.5 × 10 4 cells/well) coated with growth factor-reduced Matrigel and cultured with EGM2-MV supplemented with SEMA4D for a week. ( B ) Graph illustrating the average number of sprouts formed by DPSC cultured on 3-D collagen matrices and stimulated with 0–100 ng/mL of rhSEMA4D for 7 days. ( C ) Average number of sprouts developed by DPSC treated with 0–100 ng/mL on the 7th day. Data were analyzed from 15 microscopic fields in triplicate wells per condition and demonstrated as an average ± standard deviation. Values were compared to the control group and statistical significance was determined to be present at p < 0.05 (asterisks).

Article Snippet: After antigen retrieval, tissue sections were incubated overnight at 4 °C with rabbit anti-human SEMA4D (Bioss Inc.), mouse anti-human Plexin B1 (Santa Cruz Biotechnology Inc.) or non-specific isotype-matched IgG that was used as a negative control.

Techniques: Cell Culture, Standard Deviation

Journal: Dentistry Journal

Article Title: Semaphorin 4D Induces Vasculogenic Differentiation of Dental Pulp Stem Cells

doi: 10.3390/dj11070160

Figure Lengend Snippet: Semaphorin 4D induces SHED cells to differentiate into sprout-like structures. ( A ) Photomicrographs of SHED (×100 magnification) seeded in a 12-well plate (1.5 × 10 4 cells/well) coated with growth factor-reduced Matrigel and cultured with EGM2-MV supplemented with SEMA4D for a week. ( B ) Graph demonstrating the average number of sprouts formed by SHED cultured on 3-D collagen matrices and stimulated with 0–100 ng/mL of rhSEMA4D for 7 days. ( C ) Average number of sprouts developed by SHED treated with 0–100 ng/mL on the 7th day. Data were analyzed from 15 microscopic fields in triplicate wells per condition and demonstrated as an average ± standard deviation. Values were compared to the control group and the statistical significance was determined at p < 0.05 (asterisks).

Article Snippet: After antigen retrieval, tissue sections were incubated overnight at 4 °C with rabbit anti-human SEMA4D (Bioss Inc.), mouse anti-human Plexin B1 (Santa Cruz Biotechnology Inc.) or non-specific isotype-matched IgG that was used as a negative control.

Techniques: Cell Culture, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Mapping Semaphorins and Netrins in the Pathogenesis of Human Thoracic Aortic Aneurysms

doi: 10.3390/ijms20092100

Figure Lengend Snippet: Transcript analysis and immunohistochemistry of Semaphorins and Netrins. ( a , b ) Elastin staining (Verhoeff-Van Gieson) and degradation score in non-diseased and TAA sections of size as indicated. Dashed boxes indicate magnified areas. Scale bars represent 1000 µm and 200 µm in low and high magnified images. Arrow indicate broken elastin fragments and stars indicate areas of complete elastin degradation. *** p < 0.001. Non-diseased ( n = 4), TAA < 5.5 cm ( n = 4), TAA > 5.5 cm ( n = 5) ( c ) Quantitative PCR analysis of Semaphorin4D transcripts in non-diseased ( n = 3), TAA < 5.5 cm ( n = 4), TAA > 5.5 cm ( n = 6). ( d ) Hematoxylin and Eosin (H & E) image and representative immunofluorescence staining of Semaphorin4D (red) and Dapi (blue) in non-diseased and TAA sections of human aorta. Dashed boxes indicate magnified areas of immunofluorescence staining ( e ) Quantification of Semaphorin4D immunofluorescence staining in non-diseased and TAA. ( f ) Quantitative PCR analysis of PlexinB1 transcripts in non-diseased ( n = 3), TAA < 5.5 cm ( n = 4), TAA > 5.5 cm ( n = 6). ( g ) H & E image and representative fluorescence staining of PlexinB1 (red) and Dapi (blue) in non-diseased and TAA. ( h ) Quantification of PlexinB1 immunofluorescence staining in non-diseased and TAA. ( i ) Quantitative PCR analysis of Netrin-1 transcripts in non-diseased ( n = 3), TAA < 5.5 cm ( n = 4), TAA > 5.5 cm ( n = 5). ( j ) H & E image and representative immunofluorescence staining of Netrin-1 (red) and Dapi (blue) in non-diseased and TAA sections of human aorta. Dashed boxes indicate magnified areas of immunofluorescence staining ( k ) Quantification of Netrin-1 immunofluorescence staining in non-diseased and TAA. ( l ) Quantitative PCR analysis of Netrin-3 transcripts in non-diseased ( n = 3), TAA < 5.5 cm ( n = 4), TAA > 5.5 cm ( n = 5). ( m ) H & E image and representative immunofluorescence staining of Netrin-3 (red) and Dapi (blue) in non-diseased and TAA sections of human aorta. Dashed boxes indicate magnified areas of immunofluorescence staining. ( n ) Quantification of Netrin-3 immunofluorescence staining in non-diseased and TAA. * p < 0.05 ** p < 0.01 *** p <0.001. L = lumen. Scale bars represent 500 µm on H & E images. Magnified scale bars are 50 µm.

Article Snippet: Following is the list of primary and secondary antibodies that were used: Mouse anti-human Semaphorin4D antibody (ab212275, Abcam, Cambridge, MA, USA), Rabbit anti-human Semaphorin4D antibody (bs-6965R, Bioss antibodies, Woburn, MA, USA), Rabbit anti-human PlexinB1 antibody (ab90087, Abcam), Rabbit anti-human Netrin-1 antibody (ab126729, Abcam), Rabbit anti-human Netrin-3 antibody (bs-11059R, Bioss antibodies, Woburn, MA, USA), Goat anti-rabbit antibody Alexa Fluor 568 (A11011, Invitrogen, Wilmington, DE, USA), Goat anti-mouse antibody Alexa Fluor 568 (S11004, Invitrogen), 4′,6-diamidino-2-phénylindole (Dapi, 1: 50,000 dilution; D1306, Invitrogen).

Techniques: Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction, Immunofluorescence, Fluorescence