sheep anti human primary sema4c antibody  (R&D Systems)

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C mRNA expression is altered in different human cancers. ( A ) Oncomine database analysis results of SEMA4C mRNA levels in the tumor and normal tissues in human cancers. Note: Red and blue denote upregulation and downregulation of SEMA4C in the tumor tissues, respectively. ( B ) Interactive human body map constructed using the GEPIA webserver shows the status of SEMA4C mRNA expression in the tumor (red) and normal (green) tissue samples from different cancer types based on the TCGA data.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing, Construct

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C mRNA and protein levels are upregulated in the CRC tissues. ( A ) The relative SEMA4C mRNA expression level in normal colorectal and CRC tissues from the TCGA-COAD and TCGA-READ datasets. ( B ) Representative micrographs show SEMA4C immunohistochemical staining of the 83 pairs of colon cancer and adjacent normal colon tissue samples in the tissue microarray (TMA). Note: Scale bars=200 μm; red arrow indicates strong positive SEMA4C protein expression in the colon cancer tissue; blue arrow indicates negative SEMA4C protein expression in the adjacent normal tissue. ( C ) Quantitative analysis of SEMA4C protein expression scores based on the immunohistochemical staining of the TMA with 83 pairs of colon carcinoma and adjacent normal colon tissue samples.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing, Immunohistochemical staining, Staining, Microarray

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: The association between SEMA4C mRNA levels and the clinicopathological variables in the TCGA-CRC dataset.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques:

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: The association between SEMA4C protein levels and the clinicopathological variables in the TMA cohort of colon cancer patients.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques:

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: High SEMA4C expression correlates with poor survival outcomes in CRC patients. ( A , B ) Kaplan-Meier survival curves show OS and PFS of high- and low-SEMA4C expressing CRC patients from the TCGA database. ( C , D ) Kaplan-Meier survival curves show the OS of high- and low-SEMA4C expressing CRC patients belonging to the metastatic and non-metastatic subgroups. ( E , F ) Kaplan-Meier survival curves show OS and TFS of high- and low-SEMA4C expressing CRC patients from the TMA-colon cancer cohort.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of the prognostic significance of SEMA4C mRNA expression and other clinicopathogical parameters in TCGA-CRC patients.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of the prognostic significance of SEMA4C protein expression and other clinicopathological parameters in the TMA cohort of colon cancer patients.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: Functional enrichment analysis of GO terms and KEGG pathways related to SEMA4C and 50 SEMA4C-associated genes in CRC tissues. ( A , B ) The GO terms for the enriched ( A ) biological functions, and ( B ) cellular components related to SEMA4C and 50 SEMA4C-associated genes in the CRC tissues are shown. ( C ) The enriched KEGG pathways related to SEMA4C and 50 SEMA4C-associated genes in the CRC tissues are shown.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Functional Assay

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: GSEA enrichment plots of the TCGA-CRC dataset. GSEA results show the enrichment of gene sets related to epithelial mesenchymal transition (EMT), hedgehog signaling, Wnt/β-catenin signaling, angiogenesis, TGF-β signaling, and notch signaling in CRC patients with high SEMA4C expression. Note: NES, normalized enrichment score.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C expression correlates with the expression of EMT-related genes in CRC patients. Spearman’s correlation analysis results show the association between SEMA4C levels and the expression of EMT-related genes, including N-cadherin, β-catenin, Fibronectin1, Laminin5, COL3A1, COL4A1, CTGF, TGFβ1, ZEB1, ZEB2, SNAIL1, and SNAIL2 in the CRC tissues using the GEPIA database. Note: TPM, transcripts per million; R denotes the Spearman correlation coefficient.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C overexpression correlates with the CMS4 subtype of CRC. ( A ) Box-scatter plot shows the SEMA4C mRNA level in CMS1, CMS2, CMS3 and CMS4 subtypes of CRC patient tissues in the TCGA dataset. ( B ) ROC curve analysis shows that SEMA4C expression levels accurately discriminate between CMS4 and non-CMS4 molecular subtypes of the TCGA-CRC dataset. Note: AUC: area under receiver operating characteristic, CI: confidence interval. ( C , D ) Kaplan-Meier survival curves show the OS in the high- and low- SEMA4C expressing ( C ) CMS1-3 and ( D ) CMS4 subgroups of TCGA-CRC patients. Log-rank test was used to determine the statistical differences between the groups.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Over Expression, Expressing

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C expression negatively correlates with the DNA methylation status of the SEM4AC gene in the CRC patients. ( A ) Spearman’s correlation analysis shows the relationship between DNA methylation status of the SEMA4C gene and the expression levels of SEMA4C mRNA in the CRC samples. Note: R denotes the Spearman’s correlation coefficient. ( B ) Boxplots show the methylation levels of the CpG island shore, cg07166409 (left) and cg00605777 (right) in the SEMA4C gene in CRC patients with low- and high-expression of SEMA4C. ( C ) Kaplan-Meier survival curves show the OS of TCGA-CRC patients with hypomethylation (blue line) and hypermethylation (red line) of the SEMA4C gene in the CpG island shore, cg07166409 (above) and cg00605777 (below).

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing, DNA Methylation Assay, Methylation

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C silencing decreases proliferation, migration, and invasion of the LoVo cells. ( A ) QRT-PCR analysis shows the SEMA4C mRNA levels in the NC-siRNA- or SEMA4C-siRNA-transfected LoVo cells (human colon adenocarcinoma cell line). ( B ) CCK-8 assay results show the viability of control and SEMA4C-silenced LoVo cells. ( C ) Wound-healing assay results show the migration rates of control and SEMA4C-silenced LoVo cells. Note: Scale bars=100 μm. ( D ) Transwell invasion assay results show the invasiveness of control and SEMA4C-silenced LoVo cells. Note: Scale bars=10 μm. H indicates hour. ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant.

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Migration, Quantitative RT-PCR, Transfection, CCK-8 Assay, Control, Wound Healing Assay, Transwell Invasion Assay

Journal: Aging (Albany NY)

Article Title: High SEMA4C expression promotes the epithelial-mesenchymal transition and predicts poor prognosis in colorectal carcinoma

doi: 10.18632/aging.104038

Figure Lengend Snippet: SEMA4C silencing decreases the expression of EMT markers in the LoVo cells. ( A ) QRT-PCR analysis shows the relative mRNA expression of EMT-related genes in the control and SEMA4C-silenced LoVo cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05. ( B , C ) Representative immunofluorescence images (Scale bars=25 μm) show the levels of E-cadherin (red) ( B ) and Vimentin (green) ( C ) proteins in the control and SEMA4C-silenced LoVo cells. The nuclei are stained with Hoechst (blue).

Article Snippet: The specimens were then blocked in 10% rabbit serum for 30 min and incubated overnight with the sheep anti-human primary SEMA4C antibody (#AF6125, R & D Systems, MN, USA) at 4 o C. Then, we incubated the specimens with rabbit anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (#305-035-003, Jackson ImmunoResearch, PA, USA) for 30 min at 37 o C. Then, the specimens were incubated with 3, 3’-diaminobenzidine (DAB) and counter-stained with hematoxylin.

Techniques: Expressing, Quantitative RT-PCR, Control, Immunofluorescence, Staining