Cell- Cell Transmission of VSV-G Pseudotyped Lentivector Particles
Figure 2
VSV-G pseudotyped lentivector particles do not form aggregates.
(A) GFPvpr 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) GFPvpr 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 GFPvpr 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.