Journal: Virology

Article Title: Characterization of the Epstein-Barr virus glycoprotein BMRF-2.

doi: 10.1016/j.virol.2006.09.047

Figure Lengend Snippet: Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a retroviral vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.

Article Snippet: A human embryonic kidney cell line (293T) and a retroviral packaging cell line (ProPak A.52) were purchased from American Type Culture Collection (ATCC,Manassas, Virginia).

Techniques: Glycoproteomics, Transduction, Retroviral, Plasmid Preparation, Expressing, Staining, Fluorescence, Transfection, Membrane, Isolation, Purification, Control, Western Blot

Journal: Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

Article Title: Inhibition of NF-κB Activity Enhances Sensitivity to Anticancer Drugs in Cholangiocarcinoma Cells

doi: 10.3727/096504015x14424348426071

Figure Lengend Snippet: Figure 2. Effect of DHMEQ on 5-FU-induced cell death of CCA cell lines. Cells were treated with 5-FU with or without DHMEQ for 48 h. Cells were stained with various markers, and the image acquisitions were taken in an IN Cell Analyzer 2000. (A) Cells with nuclear fragmentation were identified by the nuclei marker, Hoechst 33342. Dead cells were identified by the presence of FITC-annexin V and/or propidium iodide. (B) Quantitative analysis of dead cell number. Bars represent the mean and SD of triplicates. The data are a representative from two independent experiments. *p < 0.05: (a) versus control, (b) versus DHMEQ, (c) versus 1 µM 5-FU, (d) versus 10 µM 5-FU.

Article Snippet: Cells were incubated further at 37°C, 5% CO 2 for 48 h, and stained with H33342, PI, and annexin V (Molecular Probes, Eugene, OR, USA) diluted in culture medium for 30 min before image acquisition in an IN Cell Analyzer 2000 (GE Healthcare, UK).

Techniques: Staining, Marker, Control