Figure 2

Characterizing the Intercellular Aggregation of Lipoplexes.

Myoblast cells were transfected with an Alexa Fluor488 labelled DNA plasmid facilitated by lipofection. (A) N&B analysis 6 h after the administration of lipoplexes. A region-of-interest (ROI) analysis was performed highlighting the cell edge (circles), cytoplasm (squares) and peri-nuclear region (triangles). (B–D) Shows the type of aggregation of particles at cell edge, cytoplasm and perinuclear regions using average brightness (2B), maximum B (2C) and particle number (2D). (E) Influence of aggregation on the mobility of the DNA (n = 12 cells for ROI analysis). (F) Confocal 3D image with orthogonal views depicting the localization of the fluorescently-labelled DNA (green) in the nucleus (blue, Hoechst 33342) after 36 h. (G) N&B analysis of the nuclear localised DNA, presented through a B vs Intensity plot and selection map (monomers, 2–12 mers, 12–25 mers, 25–40 mers and 40–58 mers (red, green, blue, orange and yellow, respectively). (H) Comparison of the aggregation and particle number of the nucleus and cytoplasm (n = 6 cells for nuclear localisation). Image size 12.8 × 12.8 μm. Graphs depict mean ± se. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p < 0.0001.