primary antibodies for mta1  (Cell Signaling Technology Inc)

Journal: Nutrients

Article Title: In Vitro Anticancer Properties of Table Grape Powder Extract (GPE) in Prostate Cancer

doi: 10.3390/nu10111804

Figure Lengend Snippet: GPE inhibits MTA1 protein expression in a dose-dependent manner in DU145 and PC3M prostate cancer cells. Immunoblots of MTA1 expression in ( A ) DU145 and ( B ) PC3M cells (top panels). β-actin was used as a loading control. Western blots were repeated three times and representative blots are shown. Quantitation of immunoblot signals (lower panels). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 (one-way ANOVA) were assessed as significant differences between treated and Ctrl vehicle cells.

Article Snippet: After blocking the membranes for non-specificity, they were probed with MTA1 (1:2000), p21 (1:1000), cleaved caspase 3 (1:1000), PTEN (1:1000), Cyclin D1 (1:1000), and pAkt (1:1000) (Cell Signaling Technology, Danvers, MA, USA) primary antibodies. β-actin antibody (1:2500) (Santa Cruz, Dallas, TX, USA) was used as a loading control.

Techniques: Expressing, Western Blot, Control, Quantitation Assay

Journal: Nutrients

Article Title: In Vitro Anticancer Properties of Table Grape Powder Extract (GPE) in Prostate Cancer

doi: 10.3390/nu10111804

Figure Lengend Snippet: Effects of GPE treatment on MTA1-associated protein levels. ( A ) Immunoblots of MTA1, Cyclin D1, and PTEN expression, in DU145 cells treated with GPE. β-actin was used as a loading control. ( B ) Quantitation of immunoblot signals of MTA1-associated proteins, Cyclin D1, and PTEN, in cells treated with 150 μg/mL GPE. ( C ) Immunoblots of MTA1, Cyclin D1, pAkt, and p21 expression, in PC3M cells treated with GPE. β-actin was used as a loading control. ( D ) Quantitation of immunoblot signals of MTA1-associated proteins, Cyclin D1, pAkt, and p21, in cells treated with 150 μg/mL GPE. Western blots were repeated three times for each protein, and representative blots are shown. *** p < 0.001 and **** p < 0.0001 (one-way ANOVA) were assessed as significant differences between 150 μg/mL GPE-treated vs. Ctrl vehicle-treated cells.

Article Snippet: After blocking the membranes for non-specificity, they were probed with MTA1 (1:2000), p21 (1:1000), cleaved caspase 3 (1:1000), PTEN (1:1000), Cyclin D1 (1:1000), and pAkt (1:1000) (Cell Signaling Technology, Danvers, MA, USA) primary antibodies. β-actin antibody (1:2500) (Santa Cruz, Dallas, TX, USA) was used as a loading control.

Techniques: Western Blot, Expressing, Control, Quantitation Assay

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: MTA1 expression in HEP-2 and HaCaT cell lines. Total RNA was extracted from the cultured cell lines and reversely transcribed to cDNA as a template for RT-PCR amplification. The level of MTA1 mRNA in the HEP-2 cell line was significantly higher than that in the HaCaT cell line. MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: RT-PCR determined the effects of pcDNA3- MTA1 , control-siRNA and MTA1 -siRNA transfection on MTA1 expression at the mRNA level. pcDNA3- MTA1 and MTA1 -siRNA were transfected into the cell line with Lipofectamine 2000. Total RNA was extracted and RT-PCR was performed. pcDNA3- MTA1 transfection increased MTA1 mRNA levels, while MTA1 -siRNA decreased MTA1 mRNA levels. MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Transfection, Expressing

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: Western blot analysis of MTA1 expression after transfection with pcDNA3- MTA1 and pSilencer3.1- MTA1 -siRNA. pcDNA3- MTA1 and MTA1 -siRNA were transfected into the cell line with Lipofectamine 2000. Whole protein was extracted and loaded for SDS-PAGE separation. The protein was transferred to a nitrocellulose membrane and pcDNA3- MTA1 increased MTA1 expression and MTA1 -siRNA greatly decreased MTA1 levels. MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Western Blot, Expressing, Transfection, SDS Page, Membrane

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: Migration ability of HEP-2 cells after MTA1 overexpression and RNAi. Cells treated by MTA1 overexpression and MTA1 silencing were loaded into the upper wells of the chamber. Cells moved to the lower side of the fibronectin-precoated polycarbonate membrane were counted and analyzed. Compared with the control, pcDNA3- MTA1 transfection promoted cell migration (P<0.05), while MTA1 -siRNA transfection suppressed cell migration (P<0.05). MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Migration, Over Expression, Membrane, Control, Transfection

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: Invasion assay of HEP-2 cells after MTA1 overexpression and RNAi. The model used in this experiment was the same as that used in the migration assay. The difference is that the membrane used in this experiment was coated with extracellular matrix Matrigel (250 μg/ml). Compared with the control, the number of cells that invaded through the membrane in the pcDNA3- MTA1 transfection group was markedly higher, while that in the MTA1 -siRNA transfection group was lower (P<0.05). MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Invasion Assay, Over Expression, Migration, Membrane, Control, Transfection

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: Adhesion assay of HEP-2 cells after MTA1 expression and RNAi. Variously treated cells were seeded into Matrigel-coated 96-well plates. The plates were washed evenly for the indicated times. The attached cells were determined to reflect their adhesion ability. Compared with the control, pcDNA3- MTA1 transfection promoted the adhesion of cells, while MTA1 -siRNA transfection suppressed the adhesion of cells (both P<0.05, compared with the control). MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Cell Adhesion Assay, Expressing, Control, Transfection

Journal: Oncology Letters

Article Title: Metastasis-associated gene 1 promotes invasion and migration potential of laryngeal squamous cell carcinoma cells

doi: 10.3892/ol.2013.1729

Figure Lengend Snippet: MTA1 overexpression and RNAi changed the wound healing ability of the HEP-2 cell line. The variously treated cells were seeded into 35 mm dishes at the same density. The wound line was drawn in the cell layer with a sterile tip. The wound healing of cells differs corresponding to pcDNA3- MTA1 or MTA1 -siRNA transfection. At the designated time points, the healing of MTA1 -siRNA transfected cells was significantly poorer. MTA1 , metastasis-associated gene 1.

Article Snippet: Blots were first incubated with MTA1 primary antibody (1:200) (Santa Cruz Biotechnology, Inc.) for 2 h at 37°C, and then with corresponding horseradish peroxidase-conjugated secondary antibody (1:2,000, Zhongshan Biotech) for 1 h at room temperature.

Techniques: Over Expression, Sterility, Transfection