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. Author manuscript; available in PMC: 2016 Jul 21.
Published in final edited form as: Nature. 2016 Jan 11;529(7586):403–407. doi: 10.1038/nature16480

Figure 3. Competence for PGCLCs versus reversion to ESCs.

Figure 3

a, Experimental design for b, c.

b, Brightfield/GFP depicting D1/D2 EpiLCs in 2i/LIF+Nanog (+Dox); note D1 EpiLCs revert to ESCs, and attempted PGCLC induction in D1 EpiLCs also results in ESC reversion (see c). Scale bar, 200 μm.

c, qPCR analysis; ΔΔCt +/− s.d. (n=2 technical replicates each from 2 biological replicates); ESCs as reference for p-values (two-sided/unpaired t-test): **p<0.01; *p<0.05.

d, Western blot for NANOG, SOX2 and α-TUBULIN (α-TUB) with ESCs as depicted; +/−Dex/Dox for 2 days. Experimental design in Extended Data Fig. 7b; gel source data in Supplementary Fig. 1.

e, Analysis (qPCR) after Sox2-KO (+Dex) and Nanog induction (+Dox); ΔΔCt +/− s.d. (n=2 technical replicates each from 2 biological replicates); parental ESCs as reference for p-values (two-sided/unpaired t-test): **p<0.01; *p<0.05. Experimental design in Extended Data Fig. 7e.

f, PGCLC induction with Dox-inducible transgenes (Nanog, Sox2 or Nanog/Sox2); D4 brightfield/GOF-GFP+ve cells (%) after FACS; scale bar, 200 μm.