Fig 1: Expression of trilineage differentiation markers. A) Matched iPSCs and differentiated endoderm, mesoderm, ectoderm cells were analyzed by immunostaining followed by flow cytometry for expression of markers specific for each germ layer (mean ± SEM, n = 7–10 cell lines). See also Fig. S1 for isotype control data, Fig. S3 for morphological changes after differentiation, Fig. S4 for paired data, and Fig. S5 for independent validation of iPSC NES staining. B) Representative flow cytometry plots showing co-immunostaining of pluripotency marker POU5F1 with each of the four candidate ectoderm markers in undifferentiated iPSCs. See also Fig. S6 for complete dataset. C, D) Expression of pluripotent marker POU5F1 (C) and four candidate ectoderm markers (D) quantified by qPCR in matched iPSCs and ectoderm differentiated cells (mean ± SEM, n = 8). Gene expression is shown as fold change of ectoderm differentiated cells compared to iPSCs, with dotted line indicating fold change of 1. * P < .05, *** P < .001, **** P < .0001, two-tailed paired t-tests.