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. 2017 May 8;7:1549. doi: 10.1038/s41598-017-01684-6

Figure 1.

Figure 1

Design and generation of XLone transgenic hPSCs. (a) Schematic showing transposable plasmid cassette design. The ColE1 origin of replication and ampicillin resistance gene for E.coli amplification are not shown. 5′ PB, 5′ PiggyBac Terminal Repeat; 3’ PB, 3’ PiggyBac Terminal Repeat; MCS, Multiple Cloning Sites; Bsdr, Blasticidin resistance gene. (b) The tight control of gene expression (GFP) is shown to have less than 1% GFP positive cells without the addition of Dox. Flow cytometry analysis of the cells after Bsd drug selection and FACS live cell sort. Live cell sorting enriched the population to maximal 70.5% GFP positive. (c) GFP positive cells with immunostaining of transcription factor pluripotency markers Oct4 and Nanog. Scale bars are 50 µm. (d) GFP positive cells with immunostaining of cell surface pluripotency marker SSEA-4. Scale bars are 100 µm.