FIG. 6.

Fluorescence reporter expression is mainly preserved during differentiation in a triple-transgenic hiPSC line. (A) Stable expression of cytoplasmic eGFP and nuclear RedStar in undifferentiated cells of the triple-transgenic line hCBiPS2 αMHCneoPGKhygro_CAG-eGFP-nRedStar6 (CAG-eGFP-nRedStar6). Depicted is a phase-contrast (left) and fluorescence image (right) of a representative colony in passage 8 after clone picking. Scale bars: 100 μm. (B) Fluorescence images of differentiated cells from the same line on day 14 of differentiation. Although both fluorescence reporters were still coexpressed in the vast majority of the differentiated cells, a minority of cells lost either eGFP or nRedStar expression (white arrows). The graph in (C) depicts the flow cytometry-based quantification of single- and double-positive cells in the pluripotent state (top) and at different time points after induction of differentiation. Color images available online at www.liebertpub.com/hgtb