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. Author manuscript; available in PMC: 2023 Jan 1.
Published in final edited form as: Methods Mol Biol. 2022;2416:13–28. doi: 10.1007/978-1-0716-1908-7_2

Figure 2. Fluorescent reporter systems to monitor the kinetics of naïve induction in live hESCs.

Figure 2.

A. Schematic of the OCT4-ΔPE-GFP reporter allele [9]: TALENs were used to disrupt the primed-specific proximal enhancer from an OCT4-2A-GFP reporter allele in WIBR3 hESCs. GFP activity is reduced in primed conditions, but stimulated upon naïve induction in 5i/L/A.

B. Phase contrast and fluorescent images of primed WIBR3 OCT4-ΔPE-GFP hESCs and naïve cells at P1 post-induction.

C. Flow cytometry analysis of OCT4-ΔPE-GFP activity in primed and naïve hESCs.

D. Schematic of a dual color reporter system targeted to both alleles of an X-linked gene (MECP2) that reports on the status of the X chromosome in female cells (MECP2-GFP/tdTomato). In the example shown, only the GFP allele is active in the primed state, but both fluorophores become expressed upon naïve induction.

E. Flow cytometry analysis of WIBR2 MECP2GFP-ON/Tom-OFF primed cells in primed hPSC medium and after conversion to the naïve state in 5i/L/A. This figure is adapted from our prior study [16].