Figure 3.
Induction of FBs into iENPs by the Seven TFs Selected Using the SOX1:EGFP Neural Reporter
(A) Schematic depiction of the experimental strategy for reducing the number of TFs (from 25 to 7) for iENP induction using the neural reporter SOX1:EGFP.
(B and C) Stepwise selection of potent iENP factors for iENP-7F generation by single TF dropouts from the original 25-TF set (B) and 13-TF set (C). The results are expressed as the relative percentage of SOX1:EGFP+ cells after each TF was removed from the TF combination.
(D) Comparison of the efficiency of induction of SOX1:EGFP+ cells from FBs by the 25-, 13-, and 7-TF combinations.
(E) Global gene expression heatmap of FBs, hESC-ENP, iENP-7F, and −13F, as determined by microarray analysis.
(F) ICC staining of iENPs-7F using antibodies against the indicated NP markers.
(G) RT-PCR analysis of endogenous and exogenous expression of the seven TFs using mRNA isolated from iENP-7F.
(H) RT-PCR analysis of the indicated neural genes using mRNA isolated from iENP-7F.
FB, fibroblasts; NC, negative control (H2O); plasmid: expression plasmids for the indicated genes. All quantitative data were obtained from three independent experiments and are presented as means ±SD. ∗p < 0.05. See also Figures S2–S4.