rabbit anti sema4d  (Santa Cruz Biotechnology)

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: Participation of Semaphorin Family and Plexins in the Clinical Course of Patients with Inflammatory Bowel Disease

doi: 10.3390/ijms252212442

Figure Lengend Snippet: Semaphorins gene expression analysis in UC patients and controls. ( A ) SEMA4D gene expression, ( B ) SEMA5A gene expression, ( C ) SEMA6D gene expression. Bars show means ± S.E.M. of SEMAS transcript levels with GAPDH as the housekeeping gene. Differences among groups were assessed by the Kruskal–Wallis test. p -value < 0.05 was considered significant.

Article Snippet: Nonspecific staining was avoided by incubating the tissues with a sniper-blocking solution (Biocare Medical, Pacheco, CA, USA) for 20 min. Tissues were incubated for 30 min at room temperature with rabbit polyclonal anti-MPO/anti-mouse SEMA4D, anti-MPO/mouse anti-PLXNC1, or mouse anti-PLXNB1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a concentration of 10 μg/mL.

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Participation of Semaphorin Family and Plexins in the Clinical Course of Patients with Inflammatory Bowel Disease

doi: 10.3390/ijms252212442

Figure Lengend Snippet: Representative image of SEMA4D/MPO detection and localization in intestinal tissue from UC and CD patients and controls without intestinal inflammation. SEMA4D-positive cells were detected in pink and MPO in brown. Burgundy arrows: Double positive cells; Red outlined arrows: SEMAD4 positive cells; Brown outlined arrows: MPO positive cells. The original magnification was 600X. (Scale bar = 50 µm).

Article Snippet: Nonspecific staining was avoided by incubating the tissues with a sniper-blocking solution (Biocare Medical, Pacheco, CA, USA) for 20 min. Tissues were incubated for 30 min at room temperature with rabbit polyclonal anti-MPO/anti-mouse SEMA4D, anti-MPO/mouse anti-PLXNC1, or mouse anti-PLXNB1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a concentration of 10 μg/mL.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Participation of Semaphorin Family and Plexins in the Clinical Course of Patients with Inflammatory Bowel Disease

doi: 10.3390/ijms252212442

Figure Lengend Snippet: Characteristics of the primers used in real-time PCR.

Article Snippet: Nonspecific staining was avoided by incubating the tissues with a sniper-blocking solution (Biocare Medical, Pacheco, CA, USA) for 20 min. Tissues were incubated for 30 min at room temperature with rabbit polyclonal anti-MPO/anti-mouse SEMA4D, anti-MPO/mouse anti-PLXNC1, or mouse anti-PLXNB1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a concentration of 10 μg/mL.

Techniques:

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Expression of Sema4D correlated with expression of HIF-1α in both HUVECs and Caco-2 cells. HUVECs and Caco-2 cells were cultured in medium containing CoCl 2 in incubator (5%CO 2 , 1%O 2 , 94%N 2 ) . The expression of HIF-1α increased in both HUVECs and Caco-2 cells after 360 minutes in a hypoxic environment. In addition, the expression level of Sema4D correlated with the expression of HIF-1α in both HUVECs and Caco-2 cells. β-actin was used as the loading control. Expression of Sema4D and HIF-1α was analyzed by NIH image software (pixel intensity of scanning) relative to β-actin controls.

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Expressing, Cell Culture, Software

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Chromatin immunoprecipitation assay of Sema4D 5' non-coding region HREs. HUVECs and Caco-2 cells were cultured under hypoxia condition (medium containing CoCl2 or cultured in an anoxic box containing GENbag). Chromatin lysates from these cells were immunoprecipitated with anti-HIF-1 antibodies according to the manufacturer's instructions. Then purified DNA was subjected to PCR with primers pairs HRE1/2 (due to their close proximity), HRE 3 and HRE 4. The VEGF promoter was used as positive control (HIF-1α). HIF-1α could bind with HRE1, HRE2, HRE4 (+) but did not bind with HRE 3 (-) in both HUVEC (A) and Caco-2 cells (B). IgG was set as negative control. Size markers are shown on the left.

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Chromatin Immunoprecipitation, Cell Culture, Immunoprecipitation, Purification, Positive Control, Negative Control

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Cloning of the Sema4D 5' non-coding region which containing mutant sites. All four specific HREs (RCGTG) were mutated (RCCAG) by overlap PCR. The Sema4D 5' UTR with site mutations were cloned into the pGL3-Basic vector.

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Clone Assay, Mutagenesis, Plasmid Preparation

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Luciferase assays of the Sema4D 5' non-coding region containing mutant HREs. HUVECs and Caco-2 cells were cultured in medium containing 0.2 mM CoCl 2 and infected with luciferase reporter vectors containing five mutation combinations. A, The Sema4D 5' non-coding region containing mutated HRE4 resulted in a significant decrease of luciferase in HUVECs. Mutation of HRE1, HRE2, HRE3 resulted in a decrease ranging from 10- 40% (A). B, Sema4D 5' untranslated region containing mutated HRE2 resulted in a significant decrease of luciferase in Caco-2 cells (B). Sample scores were analyzed by the Kruskal-Wallis Test. (* P<0.05, ** P<0.01, compared to the normal group, bars indicate mean of three individual experiments ± standard error).

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Luciferase, Mutagenesis, Cell Culture, Infection

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Mutation on Sema4D 5' non-coding region led to DNA secondary structure change. The Sema4D 5'non-coding region of ten cancer cell lines A549, Caco-2, CNE, RD, Tca-8113, SK-OV-3, Jurkat, HepG-2, SK-N-SH, U937, HL-60 were sequenced and aligned. Two distinct site mutations (T471C/C862T) of the Sema4D 5' non-coding region were detected in 7 cancer cell lines. The A600G mutation was detected in 2 cancer cell lines. These three mutations caused significant changes in the DNA secondary structure. The T471C point mutation caused fewer DNA rings between 390G-400G and loosened the tight structure between 450T-510C. The mutation at C862T exposed the retraction structure between 810C-830T, while C862T increased the distance between 665T-720T.

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Mutagenesis

Journal: Journal of Cancer

Article Title: Regulatory sequence analysis of semaphorin 4D 5' non-coding region

doi: 10.7150/jca.28169

Figure Lengend Snippet: Mutations of the Sema4D promoter affect the promoting efficiency. Four kinds of luciferase reporter plasmids containing four combinations of site mutations were infected into Caco-2 cells and HUVECs, including 1(T471C/A600A/C862C), 2 (T471T/A600A/C862T), 3 (T471C/A600A/C862T) and 4 (T471T/A600G/C862C). The normal Sema4D 5' un-coding region (T471T/A600A/C862C) acted as the control. Cells were cultured in either normal medium or medium containing 0.2 mM CoCl 2 . Luciferase reporter plasmids containing a combination of 1 and 3 promoted the expression of luciferase when compared to the control in both HUVECs (A) and Caco-2 cells (B). The sample score was analyzed by the Kruskal-Wallis Test. (* P<0.05, ** P<0.01, compared to the normal group, bars indicate mean of three individual experiments ± standard error).

Article Snippet: The primary antibodies used were as follows: rabbit anti-Sema4D (610670, BD Biosciences, 150 kDa); mouse anti-HIF-1α (D43B5, Cell Signaling Technology, 120 kDa), rabbit anti-β-actin (P30002, Abmart, 42 kDa).

Techniques: Luciferase, Infection, Cell Culture, Expressing