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Figure 3 Subsequent transfection of eGFP mRNA single-lipoplexes and mCherry mRNA single-lipoplexes. A1,B1) Transfection scheme of cells, A2,B2) representative confocal images of cells after 1 time (left panel) and 2 times (right panel) transfection with single-lipoplexes. A3,B3) Representative flow cytometry dot plots of cells receiving two subsequent transfections and A4,B4) correlation between red and green fluorescence as derived from flow cytometry dot plots. Scale bar: 100 µm. All the experiments have been triplicated in three independent days (n = 9). The mRNA dose was 0.1 µg/well per transfection round, leading to a total mRNA amount of 0.2 µg/well after the second transfection.