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. 2017 Jan 6;7:40360. doi: 10.1038/srep40360

Figure 4. Two population model to evaluate spread of the viral proteins and genome via TNTs.

Figure 4

(a) Schematics of the two population model design and its application in tracking the fate of the recipient (uninfected) cells. Cells were sorted 6 h after infection with IAV expressing the NS1-GFP construct, mixed with an equal number of RFP-A459 cells, and co-incubated at 37 °C for an additional 4–6 h. The co-incubated cells were then sorted into populations expressing detectable RFP only (R-quadrant), GFP only (G-quadrant), or RFP plus GFP (i.e. initially-uninfected RFP-A549 cells that also expressed NS1-GFP) (R/G quadrant). Cells in these three quadrants were subjected to further analysis as described in the schematic. (b) Detection of endogenous NS1-GFP transfer in A549 cells through TNTs. NS1-GFP signal is seen as bright puncta in TNTs of A549 cells infected with NS1-GFP virus or in the TNTs connecting uninfected DiD-labelled-A549 cells (red) and NS1-GFP virus infected cells (green).