Figure 3: PFV intasomes search a target DNA by 1D diffusion.
From: Retroviral intasomes search for a target DNA by 1D diffusion which rarely results in integration
(a) Experimental system for PFV intasome interaction with a target DNA using smTIRF imaging. The total internal reflection fluorescence (TIRF) field extends 200–250 nm into the flow cell (teal background). A target DNA is stretched by hydrodynamic flow and bound at both ends by the biotin–NeutrAvidin over the surface. The PFV intasomes were reconstituted with Cy3-labelled viral U5 cDNA. When the Cy3-labelled PFV intasome interacts with the target DNA inside the TIRF field, the fluorophore is excited allowing visual tracking of particles. (b) Representative trace of PFV intasome diffusion (yellow) on a target DNA (red) at a 300-ms frame rate. (c) Distribution of lifetimes (±s.e.) for wild type PFV intasomes at different NaCl salt concentrations (Supplementary Fig. 6). (d) Box plots of the diffusion coefficients distribution for wild type PFV intasomes at different NaCl salt concentrations at a 100-ms frame rate (Supplementary Fig. 6). Box plots show the mean (indentation), median (black line), upper and lower quartile (box ends), and outliers (whiskers).
