Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: Expression and location of TRAF4 in breast cancer cells. (A) Results of immunofluorescence staining showing TRAF4 (green fluorescence) localized in the cytoplasm and nuclei (scale bar, 20 μm). (B) MCF-10A, MCF-7 and MDA-MB-231 cells were lysed and the lysates were subjected to western blot analyses of TRAF4 and β-actin. Results showed that the total expression of TRAF4 in the MCF-10A cells was lower than in the MCF-7 and MDA-MB-231 cells ( ** P=0.002 and P=0.001, respectively). TRAF4, tumor necrosis factor receptor-associated factor 4; IDV, integated optical density value.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Western Blot

Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: TRAF4 suppresses the activation of NF-κB in MCF-7 Cells. The MCF-7 cells were lysed and the lysates were subjected to western blot analyses of NF-κB, β-actin and lamin B1. Following TNF-α treatment (10 ng/ml; 15 min), the nuclear expression of NF-κB was significantly downregulated when TRAF4 was knocked down in the MCF-7 cells ( ** P=0.002). TRAF4, tumor necrosis factor receptor-associated factor 4; NF-κB, nuclear factor κB.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: Biological function of TRAF4 in MCF-7 cells. (A) MTT results showing that cell proliferation was significantly suppressed following the knockdown of TRAF4 ( * P=0.02). Results of flow cytometry showed that TRAF4 ablation resulted in (B) significant increase of G 1 phase cells ( ** P=0.009) and reduction of S phase cells ( * P=0.04) and (C) significant increase in the early apoptotic cells ( ** P=0.001). All comparisons were made between the groups of MCF-7 cells or cells transfected with negative control alone. TRAF4, tumor necrosis factor receptor-associated factor 4.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Flow Cytometry, Transfection, Negative Control