Figure 5.

One pot integration of payload library. (A) Architecture of the payload library. The first fluorescent reporter (EYFP, mKate2 or Cerulean) is fused to a nuclear localization tag. The second fluorescent reporter, different from the first one, is tagged either with an utrophin localization signal, a mitochondrial localization signal or with no localization signal. All possible combinations result in a library of 18 different payloads. (B) Fluorescent microscopy images of microcolonies 7 days after selection with puromycin (10 days after transfection of equimolar amount of the 18 plasmids and BxB1 expression plasmid in HEK293FTLP chassis cell line) and no cell passaging. All 18 phenotypes were observed and are shown. (C and D) Fluorescence microscopy images of polyclonal population 14 days after transfection, with 10x (C) and 40x (D) magnification, after cell passaging. (E) Average number of each cell phenotype appearance in 500 classified cells. Error bars represent the standard deviation from these classifications performed over three different fields of views from three independently repeated experiments (total of 4500 classified cells).