a, Diagram indicating mutations observed in the two TFAP2C−/− deficient lines used in this paper. Note that both mutations cause frame shifts. b, Control and TFAP2C−/− lines are karyotypically normal. c,d,c, Expression of OCT4 (c), NANOG (d), SOX2 (e) in control and TFAP2C−/− hESCs cultured in primed conditions. The OCT4 blot is representative of 3 independent experiments. f, TRA-1-85 positive (human) cells were gated and stained for the pluripotency markers TRA-1-61 and TRA-1-80. Data represent 1 out of 3 independent experiments with similar results. g, Expression of lineage markers in primed hESCs and in embryoid bodies formed from control and TFAP2C−/− primed hESCs (n = 4 biological replicates for primed, WT EB and TFAPC2−/− EB) Mean +/− standard error is shown. Note loss of pluripotency markers upon EB differentiation and gain of neural and non-neural ectoderm markers, with a neural bias in TFAP2C−/−. Uncropped Western blot images are available in Supplementary Fig. 9. Source data for g is in Supplementary Table 8.