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

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

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Comparison of SEMA4D expression between melanoma and normal tissues. *P<0.05. Purple, melanoma; Blue, normal. SEMA4D, semaphorin 4D. SKCM, skin cutaneous melanoma; T, tumor; N, normal.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques: Comparison, Expressing

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Relevant characteristics of patients with melanoma.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Relationship between the characteristics of patients with melanoma and classification of the patients' prognosis.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Association between melanoma-related parameters and prognosis based on univariate logistic regression analysis.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Relationship between melanoma-related parameters and patient prognosis by multivariate logistic regression analysis.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Univariate Cox regression analysis of melanoma-related characteristics on the survival time of patients.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Influence of melanoma-related characteristics on patient survival time based on multivariate Cox regression analysis.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: ROC curves. ROC curves of (A) Sex, (B) Age, (C) Tumor size, (D) Family history, (E) Tumor grade, (F) SEMA4D, and (G) Tumor stage. SEMA4D, semaphorin 4D; ROC, receiver operating characteristic; AUC, area under the curve.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Receiver operating characteristic curve of all relevant factors for melanoma. SEMA4D, semaphorin 4D.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Survival curve analysis. Survival curve based on (A) Sex, (B) Age, (C) Tumor size, (D) Family history, (E) Tumor grade, (F) SEMA4D, and (G) Tumor stage. SEMA4D, semaphorin 4D; HR, hazard ratio; CI, confidence interval.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques:

Journal: Oncology Letters

Article Title: Low expression of SEMA4D as a potential predictive molecular marker of poor survival in patients with melanoma combined with liver cancer

doi: 10.3892/ol.2023.13746

Figure Lengend Snippet: Expression of SEMA4D in liver cancer tissues. (A) mRNA expression of SEMA4D. (B) Protein expression levels of SEMA4D and SEMA3B. SEMA, semaphoring; CON, control.

Article Snippet: The following antibodies were used: anti-Actin antibody (1:20,000; cat. no. 66009-1-Ig; ProteinTech Group, Inc.), anti-SEMA4D antibody (1:20,000; cat. no. 66582-1-Ig; ProteinTech Group, Inc.), anti-SEMA3B antibody (1:5,000; cat. no. ab48197; Abcam).

Techniques: Expressing, Control