Journal: Molecular Medicine Reports
Article Title: CagA promotes proliferation and inhibits apoptosis of GES-1 cells by upregulating TRAF1/4-1BB
doi: 10.3892/mmr.2017.6757
Figure Lengend Snippet: Ectopic expression of CagA increases the expression of TRAF1 and 4-1BB in GES-1 cells. (A) GES-1 cells were transfected with plasmid pEGFP-C1/CagA or control vector pEGFP-C1 for 48 h. The mRNA level of TRAF1 was determined using RT-qPCR analysis with GAPDH as the internal control. (B) GES-1 cells were transfected with plasmid pEGFP-C1/CagA or control vector pEGFP-C1 for 48 h. The mRNA level of 4-1BB was determined using RT-qPCR analysis with GAPDH as the internal control. (C) GES-1 cells were transfected with plasmid pEGFP-C1/CagA or control vector pEGFP-C1 for 48 and 72 h. TRAF1 and 4-1BB protein levels were determined using western blot analysis with GAPDH as the loading control. (D) Relative expression of TRAF1 compared with GAPDH. (E) Relative expression of 4-1BB compared with GAPDH. *P<0.05, **P<0.01, ***P<0.001. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; EGFP, enhanced green fluorescent protein; CagA, cytotoxin-associated gene A; TRAF1, tumor necrosis factor receptor-associated factor 1; 4-1BB, tumor necrosis factor receptor superfamily member 9.
Article Snippet: Rabbit anti-CD137 polyclonal antibody was purchased from Abcam (Cambridge, UK; cat. no. ab203391); rabbit anti-TRAF1 (45D3) monoclonal antibody was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA; cat. no. 4715); rabbit anti-CagA (cat. no. sc-25766), goat anti-rabbit immunoglobulin (Ig)G-horseradish peroxidase (HRP; cat. no. sc-2030) and goat anti-mouse IgG-HRP (cat. no. sc-2302) antibodies were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA); and mouse anti-GAPDH monoclonal antibody (cat. no. MAB374) was purchased from Merck KGaA (Darmstadt, Germany).
Techniques: Expressing, Transfection, Plasmid Preparation, Control, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction
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