Journal: PLoS ONE
Article Title: Cell- Cell Transmission of VSV-G Pseudotyped Lentivector Particles
doi: 10.1371/journal.pone.0074925
Figure Lengend Snippet: ( A ) GFP vpr labeled vector particles were allowed to settle on glass slides in the absence (top) or presence (bottom) of recombinant fibronectin fragment, CH296, stained with anti-VSV-G antibody and Alexa Fluor 647 anti-mouse secondary antibody, and visualized via deconvolution microscopy. ( B ) Particles either counted as “individual” (gray bars) or as “aggregate” (black bars) if 2 or more individual particles were observed to occupy contiguous space. Significance was determined by performing a Student’s 2-tailed t-test , assuming unequal variance between groups. ( C ) GFP vpr labeled particles were allowed to settle on glass slides in the absence (top) or presence (bottom) of 8 µg/ml protamine sulfate. ( D ) Particles either counted as “individual” (gray bars) or as “aggregate” (black bars) if 2 or more individual particles were observed to occupy contiguous space. Asterisk indicates that data set for “(-PS) control is the same from (B), “(-FN)” control. Significance was determined by performing a Student’s 2-tailed t-test , assuming unequal variance between groups. ( E ) Jurkat (top panels) and 293T cells (bottom panels) were exposed to GFP vpr vector particles (green) for 1 hour at 37C, washed, and fixed with 4% paraformaldehyde. Cells were stained with phalloidin (red) and anti-VSV-G antibody (magenta), followed by staining with anti-rabbit Alexa Fluor 647 (blue), and imaging via deconvolution microscopy. ( F ) Particles either counted as “individual” (gray bars) or as “aggregate” (black bars) if 2 or more individual particles were observed to occupy contiguous space. Significance was determined by performing a Student’s 2-tailed t-test , assuming unequal variance between groups.
Article Snippet: Experiments involving fibronectin fragment CH296 (Takara Mirus, Madison, WI) used a final concentration of 5 µg/ml, unless otherwise noted.
Techniques: Labeling, Plasmid Preparation, Recombinant, Staining, Microscopy, Imaging
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