Journal: Journal of Virology

Article Title: The Conserved YAGL Motif in Human Metapneumovirus Is Required for Higher-Order Cellular Assemblies of the Matrix Protein and for Virion Production

doi: 10.1128/jvi.02694-10

Figure Lengend Snippet: FIG. 6. Immunofluorescence analysis of the intracellular distributions of wild-type and mutant M proteins. 293T cells harboring stable replicons of the indicated hMPV-GFP-Puro clones (A and B) or Vero cells infected with the hMPV-GFP-Puro-wt or FAGL clone (C) were grown on glass coverslips and processed for immunofluorescence analysis using anti-M antibodies (red). The nuclei were stained with DAPI (blue). In panel A, the top row shows low-magnification images of the cells, whereas the bottom row presents higher-magnification images of the areas defined by the insets in the upper panels. “293T” and “Vero” represent naïve cells. Images were acquired using a motorized spinning-disc confocal microscope (Yokogawa CSU-22 confocal head and Zeiss Axiovert 200 M microscope). Maximum-intensity projections of confocal z stacks are depicted in panels A, B, and C. (D) 3D renditions (generated with the 3D volume view function of SlideBook software) of cells grown and processed as in panels A and B. Bars and grids, 10 m.

Article Snippet: Images were acquired using a motorized spinning-disc confocal microscope (Yokogawa CSU-22 confocal head and Zeiss Axiovert 200 M microscope) and were analyzed with SlideBook software (version 4.2.0; Intelligent Imaging Innovations).

Techniques: Mutagenesis, Clone Assay, Infection, Staining, Microscopy, Generated, Software

Journal: Journal of Virology

Article Title: The Conserved YAGL Motif in Human Metapneumovirus Is Required for Higher-Order Cellular Assemblies of the Matrix Protein and for Virion Production

doi: 10.1128/jvi.02694-10

Figure Lengend Snippet: FIG. 7. Confocal fluorescence microscopy of cellular wild-type and mutant M proteins fused to GFP. 293T cells were transfected with plasmids expressing the wild-type (wt) M protein or the indicated M mutants, all fused to GFP. The cells were fixed, and their nuclei were stained with Hoechst dye (blue). (A) Low-magnification images of the cultures. (B) Higher-magnification images of the areas defined by the insets in panel A. (C) Confocal images of single cells of the indicated transfected cultures. Orthogonal images are presented above and to the left of each regular imaging plane (each central panel is an x-y image, the top panel is an x-z image, and the left panel is a z-y image). Images were acquired using a motorized spinning-disc confocal microscope (Yokogawa CSU-22 confocal head and Zeiss Axiovert 200 M microscope). Bars, 10 m. GFP, free GFP that is not fused to the M protein.

Article Snippet: Images were acquired using a motorized spinning-disc confocal microscope (Yokogawa CSU-22 confocal head and Zeiss Axiovert 200 M microscope) and were analyzed with SlideBook software (version 4.2.0; Intelligent Imaging Innovations).

Techniques: Microscopy, Mutagenesis, Transfection, Expressing, Staining, Imaging

Journal:

Article Title: Ovarian Cancer Cell Detachment and Multicellular Aggregate Formation Are Regulated by MT1-MMP: A Potential Role in Intra-Peritoneal Metastatic Dissemination

doi: 10.1158/0008-5472.CAN-08-4151

Figure Lengend Snippet: (A) Expression of MT1-MMP RNA in DOV13 MCAs or 2D cultures. Real time RT-PCR was performed to detect RNA expression. Shown is the average of three independent experiments ± standard deviation. (*p<.05). (B) Expression of MT1-MMP protein. Western blot was performed to detect relative MT1-MMP protein levels with β-tubulin as a loading control. Arrow - 55 kDa active MT1-MMP; arrowhead - 43 kDa MT1-MMP autolysis product. (C) Representative MCA formed by OVCA 433 or DOV13 cells. (D) Expression of MT1-MMP augments MCA formation. Cells were cultured for MCA formation using the hanging drop method and measured using a Zeiss Axiovert Imaging Software. Data shown are expressed in μm, and represent the mean diameter measurement of n=10 MCAs. (+) indicates p<.05 relative to vector-transfected cells; (*) indicates p<.05 relative to cells expressing wild type MT1-MMP. Untransfected DOV13 cells, that express similar levels of endogenous MT1-MMP, are shown for comparison.

Article Snippet: Cells were cultured for MCA formation using the hanging drop method and measured using a Zeiss Axiovert Imaging Software.

Techniques: Expressing, Quantitative RT-PCR, RNA Expression, Standard Deviation, Western Blot, Cell Culture, Imaging, Software, Plasmid Preparation, Transfection